Federal State Educational Institution
secondary vocational education
"Krasnoyarsk Medical and Pharmaceutical College
Federal Agency for Healthcare and Social Development "
N.V. Vlasova
Methods
clinical laboratory research
in the field of secondary medical education as a teaching aid for students of secondary medical educational institutions,
students in the specialty 060110 "Laboratory diagnostics"
Krasnoyarsk
Reviewer: D.A. Grishchenko, chief specialist in clinical and laboratory
Health and Drug Agency Diagnostics
Provisions for the administration of the Krasnoyarsk Territory, head
Clinical and Diagnostic Laboratory of the Krasnoyarsk Territory
Hospitals # 1.
Vlasova N.V.
В 58 Methods of clinical laboratory research: Instructional
Benefit. / N.V. Vlasov. - Krasnoyarsk: Krasnoyarsk medical
College of Pharmacy, 2008.- 222s.
This tutorial is a systematic material on clinical laboratory research methods.
Consists of two sections. The first section contains information on the methods of obtaining and laboratory research of urine, gastric juice, bile, feces, cerebrospinal fluid, sputum, genital secretions, fluids of serous cavities, as well as the results of these studies in the norm and the nature of their changes in diseases. The second section of the manual is devoted to hematological studies.
It is intended for students of secondary specialized educational institutions studying in the specialty "Laboratory diagnostics".
List of abbreviations ………………………………………………………………………… .9
Foreword ………………………………………………………………………………… 10
Introduction …………………………………………………………………………………… ..11
^ SECTION I. GENERAL CLINICAL RESEARCH ….......................13
Chapter 1. Urine examination………………………………………………………………..13
Formation and composition of urine ... …………………………………………………… ... 13
Urine test …………………………………………………………………… .14
1.2.1.1. The amount of urine ……………………………………………………………… ..15
1.2.1.2. Urine color ……………………………………………………………………… ..15
1.2.1.3. Transparency of urine ………………………………………………………… ... 16
1.2.1.4. Urine reaction …………………………………………………………………… .17
1.2.1.5. Urine smell …………………………………………………………………… .18
1.2.1.6. Relative density of urine …………………………………………… ... 18
1.2.1.7. Zimnitsky's test …………………………………………………………… 18
1.2.1.8. Test questions on the topic "Research of physical
Properties of urine "………………………………………………………………… ... 20
1.2.2. Chemical analysis of urine …… …………………………………………… ..20
1.2.2.1. Determination of protein in urine ………………………………………………… ... 20
1.2.2.2. Determination of glucose in urine ……………………………………………… ..25
1.2.2.3. Determination of ketone bodies in urine …………………………………………… 27
1.2.2.4. Determination of urobilin and bilirubin in urine ………………………………. 28
1.2.2.5. Determination of blood pigment in urine …………………………………… ..30
1.2.2.6. Control questions on the topic "Chemical study of urine" ... ... ... ... 31
1.2.3. Microscopic examination of urine sediment …………………………………… ..31
1.2.3.1. Indicative method …………………………………………………… ..31
1.2.3.2. Quantitative methods ……………………………………………………… ..36
1.2.3.3. Control questions on the topic "Microscopic examination
Urine sediment "…………………………………………………………………… 38
1.2.4. Examination of urine using test strips ……………………………………… 38
1.3. Urinary syndromes ………………………………………………………………… ... 39
1.4. Final control questions to the chapter "Study of urine" ……………… 41
Chapter 2. Study of gastric secretion ……………………………………………44
2.1. Functions of the stomach. The composition of gastric juice ………………………………………… ..44
2.2. Methods for the study of gastric secretion ………………………………………… ... 45
2.2.1. Phases of gastric secretion …………………………………………………………. 45
2.2.2. Fractional method of gastric sounding …………………………………… ..46
2.2.3. Control questions on the topic "Methods for the study of gastric
Secretions "…………………………………………………………………………… 47
2.3. Study of gastric juice ... ………………………………………………… ..47
2.3.1. Physical properties ………………………………………………………………… 48
2.3.2. Chemical research ……………………………………………………… ... 48
2.3.2.1. Determination of acidity ……………………………………………………… 48
2.3.2.2. Determination of the flow rate of hydrochloric acid ……………………………………… ... 50
2.3.2.3. Determination of hydrochloric acid deficiency …………………………………… ..50
2.3.2.4. Determination of lactic acid ……………………………………………… .51
2.3.2.5. Determination of proteolytic activity ………………………………… .51
2.3.2.6. Intragastric pH-metry ………………………………………………… 52
2.3.3. Microscopic examination of gastric contents …………………… 52
2.3.4. Control questions on the topic "Study of gastric juice" ..................... 53
2.4. Probeless methods for assessing the acidity of gastric juice ………………………… 53
2.5. Final control questions for the chapter "Research
Gastric secretion "………………………………………………………………… 54
Chapter 3. Study of duodenal contents ……………………………………..56
3.1. Composition and function of bile. Physiology of formation and excretion of bile …………… ..56
3.2. Duodenal intubation methods ………………………………………………… ..57
3.3. Study of duodenal contents …………………………………………… .59
3.3.1. General properties ……………………………………………………………………… .59
3.3.2. Microscopic examination ………………………………………………… 60
3.4. The diagnostic value of duodenal intubation …………………………… ... 62
3.5. Control questions to the chapter "Study of duodenal contents" ...................... 63
Chapter 4. Stool examination …………………………………………………………………64
4.1. Stool composition ……………………………………………………………………………. 64
4.2. Stool examination ……………………………………………………………………… ... 64
4.2.1. General properties of feces ……………………………………………………………… .64
4.2.2. Chemical examination of feces …………………………………………………… 67
4.2.3. Control questions on the topic "Physical and chemical properties of feces" ...................... 68
4.2.4. Microscopic examination of feces …………………………………………… ... 69
4.2.4.1. Microscopic elements of feces ………………………………………… .69
4.2.4.2. Protein food residues in feces ………………………………………………… 70
4.2.4.3. Residues of carbohydrate food in feces ……………………………………………… 71
4.2.4.4. Fat residues in feces ……………………………………………………… ..72
4.2.4.5. Stool cell elements …………………………………………………… 73
4.2.4.6. Crystalline formations ……………………………………………… .73
4.2.4.7. Microflora ………………………………………………………………… ..73
4.2.4.8. Control questions on the topic "Microscopic examination of feces" ... 75
4.3. Scatological syndromes ………………………………………………………… ... 75
4.4. Final control questions for the chapter "Stool research" ...................... 77
Chapter 5. Cerebrospinal fluid examination …………………………………..78
5.1. Formation, functions and receipt of cerebrospinal fluid ............................................................................. 78
5.2. The study of cerebrospinal fluid …………………………………………………………………… .79
5.2.1. Physical properties of cerebrospinal fluid …………………………………………………… ..79
5.2.2. Microscopic examination of cerebrospinal fluid ………………………………………… .80
5.2.3. Chemical research of cerebrospinal fluid ……………………………………………… .82
5.3. Characteristics of cerebrospinal fluid in some diseases of the central nervous system ...................... 84
5.4. Control questions to the chapter "Research of cerebrospinal fluid" ... ... ... 86
Chapter 6. Study of exudates and transudates……………………………………….87
6.1. Types of points ……………………………………………………………………… .87
6.2. Investigation of fluids of serous cavities ............................................................. 88
6.2.1. Determination of physical and chemical properties ………………………………………… 89
6.2.2. Microscopic examination ………………………………………………… ... 89
6.3. Control questions to the chapter "Research of exudates and transudates" ..................... ... 91
Chapter 7... Sputum examination …………………………………………………………….91
7.1. Sputum collection …………………………………………………………………………. 92
7.2. Safety rules for working with sputum ……………………………… ..93
7.3. Sputum test …………………………………………………………………… 94
7.3.1. Determination of the general properties and nature of sputum ................................................................. ... 94
7.3.2. Control questions on the topic "General properties of sputum" ……………………… 97
7.3.3. Microscopic examination of sputum ………………………………………… 97
7.3.3.1. Preparation and study of native sputum preparations ………………… .97
7.3.3.2. Cellular elements of sputum ……………………………………………… 98
7.3.3.3. Fibrous formations in sputum ………………………………………… .99
7.3.3.4. Crystalline formations of sputum ... ………………………………… .100
7.3.4. Bacterioscopic examination of sputum …………………………………… .101
7.3.4.1. Preparation and fixation of smears ………………………………………… ... 101
7.3.4.2. Ziehl-Nielsen staining ……………………………………………… .102
7.3.5. Test questions on the topic "Microscopic and
Bacterioscopic examination of sputum "…………………………………… .104
7.4. Characteristics of sputum in some diseases of the respiratory system ... ... .104
7.5. Final control questions to the chapter "Sputum examination" ………… 105
Chapter 8... Examination of genital discharge …………………………………106
8.1. Laboratory tests for infections transmitted primarily
Sexually transmitted …………………………………………………………………………. 106
8.1.1. Syphilis …………………………………………………………………………… 106
8.1.2. Gonorrhea …………………………………………………………………………… .109
8.1.3. Urogenital chlamydia ……………………………………………………… ... 109
8.1.4. Urogenital trichomoniasis ……………………………………………………… 111
8.1.5. Bacterial vaginosis ………………………………………………………… ... 112
8.1.6. Urogenital candidiasis ………………………………………………………. 112
8.1.7. Control questions on the topic "Laboratory research for STIs" ....... .113
8.2. Examination of the vaginal contents …………………………………………… ... 114
8.2.1. Cytological studies ………………………………………………… .114
8.2.1.1. Sampling and preparation of preparations for microscopy ………… 114
8.2.1.2. Morphology of vaginal epithelial cells ……………………………. 115
8.2.1.3. Cytological assessment of vaginal smears ……………………………… .116
8.2.2. Determination of the degree of purity of the vaginal contents ............................................................. 118
8.2.3. Control questions on the topic "Examination of the contents of the vagina" ... ... ... 119
8.3. Examination of ejaculate and prostate secretion ............................................................. 119
8.3.1. Composition and production of seminal fluid .............................................................. 120
8.3.2. Ejaculate examination ……………………………………………………………… 121
8.3.2.1. Physical and chemical research ………………………………………… ... 121
8.3.2.2. Microscopic examination of ejaculate ………………………………… .122
8.3.3. Study of the secretion of the prostate gland …………………………………… 125
8.3.4. Control questions on the topic "Research of ejaculate and
Prostate secretion "………………………………………………. 126
Chapter 9. Laboratory diagnostics of mycoses …………………………………………...127
9.1. Classification of mycoses …………………………………………………………… ... 127
9.2. Technique for taking material and preparing preparations for
Microscopic examination ………………………………………………… .128
9.3. Laboratory diagnostics of fungal skin diseases ………………………… ... 129
9.4. Rules for safe work in a mycological laboratory ......................................................... 131
9.5. Control questions to the chapter "Laboratory diagnostics of mycoses" …………………………………………………………………………………………………………… 131 131
Section II. HEMATOLOGICAL STUDIES…………. 132
Chapter 1... General clinical blood test …………………………………………...132
Composition and functions of blood .............................................................................. 132
Taking blood for research ………………………………………………………………………………………………………………………………………………………… 133 133
Determination of the concentration of hemoglobin in the blood …………………………………… .135
1.3.2. Methods for determining the concentration of hemoglobin in the blood ……………………… ..137
1.3.3. Clinical significance of blood hemoglobin …………………………………… ..137
1.3.4. Control questions on the topic "Determination of concentration
Hemoglobin of blood "……………………………………………………………… .138
1.4. Determination of the erythrocyte sedimentation rate ………………………………………… 138
1.4.1. Factors affecting ESR ………………………………………………………. 138
1.4.2. Methods for determination of ESR ………………………………………………………. 139
1.4.3. Clinical significance of ESR …………………………………………………… ... 139
1.4.4. Control questions on the topic "Determination of ESR" …………………………… .140
1.5. Determination of the number of leukocytes in the blood ……………………………………… ... 140
1.5.1. Functions of leukocytes ……………………………………………………………. 140
1.5.2. Methods for counting the number of leukocytes in the blood ……………………………… ..141
1.5.3. The clinical significance of the number of leukocytes in the blood ………………………… .142
1.5.4. Control questions on the topic "Determination of the number of leukocytes
In the blood "…………………………………………………………………………… .143
1.6. Determination of the number of erythrocytes in the blood ……………………………………… ... 143
1.6.1. Functions of erythrocytes ……………………………………………………………… .144
1.6.2. Methods for counting the number of erythrocytes in blood ………………………………… 144
1.6.3. The clinical significance of the number of erythrocytes in the blood ......................................................... 145
1.7.1. Color index of blood …………………………………………………… .146
1.7.2. Control questions on the topic "Determination of the quantity
Red blood cells. Color index of blood "……………………………… .147
1.8. Calculation of the leukocyte formula ………………………………………………… ... 147
1.8.1. Morphology of certain types of peripheral blood leukocytes is normal ....... 147
1.8.2. Methods for calculating the leukocyte formula ……………………………………… ..149
1.8.2.1. Preparation of smears ……………………………………………………………………………………………………………………………… 149
1.8.2.2. Staining of smears …………………………………………………………… ... 150
1.8.2.3. Technique for counting the leukocyte formula ………………………………… 152
1.8.3. Leukocyte formula in normal and pathological conditions ……………………………… ..152
1.8.3.1. Leukocyte formula is normal …………………………………………… .152
1.8.3.2. Changes in the morphology of leukocytes in pathology ……………………… ... 153
1.8.3.3. Change in the number of certain types of leukocytes in pathology ... ... ... 154
1.8.4. Control questions on the topic "Calculation of the leukocyte formula" ... ... ... ... ... 155
1.9. Changes in blood in some conditions and diseases ...................... 155
1.9.1. Age features of blood ………………………………………………… .155
1.9.2. Change in blood during pregnancy …………………………………………… ..156
1.9.3. Hereditary anomalies of leukocyte morphology …………………………… ..157
1.9.4. Changes in blood with purulent-inflammatory and infectious
Diseases ………………………………………………………………………… 158
1.10. Final control questions for the chapter "General clinical
Blood test "………………………………………………………………………… ... 158
Chapter 2. Automatic methods of research of blood cells… ……………………159
Chapter 3. Hematopoiesis scheme…………………………………………………………….163
Chapter 4. Anemias……………………………………………………………………………...165
4.1. Classification of anemias ………………………………………………………………… .165
4.2. Laboratory signs of anemia …………………………………………………… ..167
4.2.1. Changes in the morphology of erythrocytes in anemia ……………………………… ..167
4.3. Anemia due to blood loss …………………………………………………… ..170
4.3.1. Acute post-hemorrhagic anemia ……………………………………………… .170
4.3.2. Chronic post-hemorrhagic anemia ……………………………………… ... 170
4.3.3. Test questions on the topics “Laboratory signs of anemia.
Anemia due to blood loss "……………………………………………… ... 170
4.4. Anemia due to impaired blood formation …………………………………… 171
4.4.1. Iron deficiency anemias ………………………………………………………… 171
4.4.2. Iron-saturated anemias ... ………………………………………………… .172
4.4.3. В 12 (folic acid) -deficient anemias .............................................................. 172
4.4.4. Hypo- and aplastic anemias ... ………………………………………………… ..173
4.4.5. Control questions on the topic "Anemia due to violation
Blood formation "………………………………………………………………… .174
4.5. Hemolytic anemias ... …………………………………………………………… ... 174
4.5.1. Causes and signs of hemolytic anemias …………………………………… 174
4.5.2. Classification of hemolytic anemias ……………………………………… ... 175
4.5.3. Hemolytic disease of newborns ……………………………………… 176
4.6. Determination of hematocrit value …………………………………………… ..177
4.7. Counting the number of reticulocytes …………………………………………………… .178
4.8. Determination of osmotic resistance of erythrocytes …………………………… 179
4.9. Final control questions for the chapter "Anemias" ………………………… ..181
Chapter 5. Radiation sickness …………………………………………………………………...182
5.1. Acute radiation sickness ……………………………………………………………… .183
5.2. Chronic radiation sickness ………………………………………………………… ..185
5.3. Control questions on the topic "Radiation sickness" ………………………………… ... 185
Chapter 6... Leukemia…………………………………………………………………………….186
6.1. Etiology, pathogenesis, classification of leukemia …………………………………………………………………………………………………………………………………… 186
6.2. Acute leukemia ……………………………………………………………………… .187
6.2.1. Classification of acute leukemia ……………………………………………… ... 187
6.2.2. Clinical manifestations and blood picture in acute leukemia ……………… ... 188
6.2.3. Cytochemical characteristics of blast cells in acute leukemia ... ... ... .190
6.2.4. Control questions on the topic "Acute leukemia" ……………………………… .191
6.3. Chronic leukemia ………………………………………………………………… 191
6.3.1. Myeloproliferative diseases …………………………………………… ... 191
6.3.1.1. Chronic myeloid leukemia ………………………………………………… .192
6.3.1.2. Erythremia …………………………………………………………………… 193
6.3.1.3. Chronic monocytic leukemia ……………………………………… .193
6.3.1.4. Control questions on the topic "Myeloproliferative diseases" ... .194
6.3.2. Lymphoproliferative diseases ………………………………………… ... 194
6.3.2.1. Chronic lymphocytic leukemia ……………………………………………… ... 195
6.3.2.2. Multiple myeloma ………………………………………………… ..196
6.3.2.3. Control questions on the topic "Lymphoproliferative
Diseases "……………………………………………………………… ... 197
6.4. Final control questions to the chapter "Leukemia" …………………… ..197
Chapter 7. Leukemoid reactions …………………………………………………………..198
Chapter 8. Hemorrhagic diathesis … …………………………………………………….200
8.1. Classification of hemorrhagic diathesis ………………………………………… .200
8.2. Determination of the number of platelets in the blood ……………………………………… ..201
8.2.1. Morphology and functions of platelets ………………………………………… ... 201
8.2.2. Methods for determining the number of platelets …………………………………… 202
8.2.3. The clinical significance of the number of blood platelets ………………………… ..203
8.3. Determination of the duration of bleeding and clotting time
Capillary blood …………………………………………………………………… 204
8.4. Control questions to the chapter "Hemorrhagic diathesis" ................................................................... 205
Chapter 9... Groups and Rh-affiliation of blood ……………………………………….205
9.1. Blood groups of the AB0 system ………………………………………………………… 206
9.1.2. Methods for determining the blood group …………………………………………… .207
9.2. Rh-belonging of blood ……………………………………………………… ..212
9.3. Control questions to the chapter "Groups and Rh-belonging of blood" ………… .214
Chapter 10... Quality control of laboratory tests …………………………...215
Standards of answers to test tasks …………………………………………………… .220
Bibliographic list ……………………………………………………………… .221
^ List of abbreviations
ACTH - pituitary adrenocorticotropic hormone
B - basophil
i / v - intravenously
i / m - intramuscularly
WHO - World Health Organization
HDN - hemolytic disease of the newborn
DNA - deoxyribonucleic acid
Duodenum - duodenum
Ischemic heart disease
IS - maturation index
STIs - Sexually Transmitted Infections
CI - karyopyknotic index
CDL - clinical diagnostic laboratory
ACM - acid-fast mycobacteria
L - lymphocyte
LB - radiation sickness
MON - monocyte
MPO - myeloperoxidase
Np / i - stab neutrophil
NS / I - segmented neutrophil
OL - acute leukemia
ARS - acute radiation sickness
ARVI - acute respiratory viral infection
s / c - subcutaneously
RNA - ribonucleic acid
SI - international system of units of measurement
SMS - synthetic detergent
ESR - erythrocyte sedimentation rate
FEK - photoelectric colorimeter
CLL - chronic radiation sickness
CML - chronic myeloid leukemia
CRF - chronic renal failure
CNS - central nervous system
CPK - color index of blood
CSF - cerebrospinal fluid
E - eosinophil
EDTA - ethylenediaminetetraacetate
EI - eosinophilic index
Foreword
The importance of laboratory research at the present stage of development of medicine is constantly increasing.
The main contingent in terms of the number of employees of clinical diagnostic laboratories is made up of laboratory assistants with secondary specialized education, which makes special requirements for their training. The lack of a sufficient number of modern textbooks on the methods of clinical laboratory research for secondary specialized educational institutions in the context of a sharp expansion of the range of laboratory research and technical re-equipment of clinical diagnostic laboratories determines the need to publish a textbook on clinical laboratory diagnostics for medical laboratory technicians.
This study guide includes two sections - general clinical and hematological studies, consisting of several chapters. Each chapter is devoted to the laboratory analysis of a certain type of biological material (urine, contents of the gastrointestinal tract, sputum, cerebrospinal fluid, genital secretions, exudative fluids, blood) and contains information on the methods of obtaining them and standardized methods of laboratory research, as well as the results of these studies in the norm and the nature of their changes in diseases.
The materials of the manual are presented in accordance with the documents regulating the activities of the clinical diagnostic laboratories of the healthcare facilities of the Russian Federation. Thus, the chapter "Quality control of clinical laboratory research" covers the modern concept of the issue in accordance with the Order of the Ministry of Health of the Russian Federation No. 45 of February 7, 2000. The topic "Sputum research" contains recommendations of Appendix No. 10 to the order of the Ministry of Health of Russia dated March 21, 2003. No. 109 "Instructions for unified methods of microscopic examinations for the detection of acid-fast mycobacteria in clinical diagnostic laboratories of health care facilities." Questions of determining the group and Rh-belonging of blood are given in accordance with the Order of the Ministry of Health of the Russian Federation No. 2 dated January 9, 1998 "On the approval of instructions for immunoserology."
At the end of each topic there are control questions, and at the end of the major chapters there are final questions in the form of tests to reconnect with the response standards at the end of the manual. The selected form allows a limited number of test items to cover a large amount of material.
The manual reflects the experience gained over many years of teaching the discipline "Methods of clinical laboratory research."
Introduction
The discipline "Methods of clinical laboratory research" studies a complex of physicochemical and biological methods used to obtain objective data on the state of the human body.
As a scientific discipline, clinical laboratory diagnostics arose at the intersection of clinical medicine, anatomy, physiology, biology, physics, chemistry and other sciences. She solves the following tasks:
Development of optimal methods for the study of biological material;
Establishment of limits for fluctuations in the norm for certain groups of people (by sex, age, habitat, etc.);
Establishing the diagnostic value of individual laboratory tests.
The main task of clinical laboratory diagnostics in practical medicine is to help the attending physician in making a diagnosis of the disease, treating patients, and taking preventive measures.
The main objects of clinical laboratory research are the contents of blood vessels and cavities (blood, cerebrospinal fluid, transudates and exudates, gastric juice, bile), excretions of the human body (urine, feces, sputum, seminal fluid), as well as bone marrow, punctates of lymph nodes, etc. ...
The composition and properties of human biological fluids have attracted the attention of scientists for a long time. So, already in the treatises of ancient India and China (X-VI centuries BC) there are indications of the study of the properties of urine. The Uzbek doctor Abu Ali ibn Sina (Avicenna) in his works associates a change in the nature of a person's secretions (urine, feces) with certain diseases. However, these observations of ancient scientists were limited only to a description of the general properties (color, quantity, smell, etc.) of biological material. The development of laboratory diagnostics as a scientific discipline was facilitated by the invention of the microscope and colorimeter, the discovery of the structure of the cell, and other advances in natural science. The first primitive clinical and diagnostic studies associated with an attempt to apply the methods of chemical analysis in medicine date back to the 16th century - the beginning of the Renaissance.
In Russia, the first clinical diagnostic laboratory was organized by the outstanding clinician S.P. Botkin at the therapeutic department of the Military Medical Academy of St. Petersburg. There are great merits in the development of laboratory work by D.L. Romanovsky, who proposed his own method for staining the formed elements of blood, which is still used today. A significant contribution to laboratory work was made by domestic scientists VE Predtechensky, MN Arinkin (method of intravital bone marrow extraction), IA Kassirsky (monograph "Clinical Hematology"), EA Kost (organized the All-Union Society of Laboratory Doctors, the journal "Laboratory Business"), etc.
In modern clinical laboratory diagnostics, methods of optical, ionometric, immunoassay, electrophoretic, chromatographic and other types of analysis, methods of "dry" chemistry are widely used. For carrying out many types of laboratory research, the production of special reagent kits has been established, which significantly improves the quality of analyzes. In many clinical diagnostic laboratories of healthcare facilities, high-tech analyzers are used to perform laboratory research in a fully automated mode.
In all laboratories, studies are carried out using uniform unified methods approved by the Ministry of Health of the Russian Federation and mandatory for all CDLs.
Special attention of laboratory service specialists is paid to improving the quality of analyzes, which is ensured by the introduction of special programs using control materials into everyday practice of the CDL.
SECTION I
^ GENERAL RESEARCH
__________________________________________________________________
Chapter 1
EXAMINATION OF URINE
FORMATION AND COMPOSITION OF URINE
Urine formation.Urine is formed in the kidneys, the main function of which is to maintain the constancy of the internal environment of the body. This function is ensured by the excretion of metabolic end products, excess salts and water, as well as toxic and foreign substances in the urine.
The urinary organs include the kidneys [lat.ren, Greek. nephros], ureters [lat.ureter], bladder [lat.cystis], urethra [lat.urethra]. The renal pelvis is located inside the kidneys.[lat. pyelos] ... The main functional unit of the kidneys is the nephron - a collection of tubular tubules with vascular glomeruli.
Urine formation occurs in 3 stages.
^ Stage 1 - filtration , during which the so-called "primary" urine is formed, which differs from blood plasma only in the absence of coarse proteins, since they do not pass through the renal filter due to the very large size of the molecules. Plasma filtration occurs in the glomeruli due to the increased blood pressure in the capillaries of the renal glomerulus, which is created due to the significantly smaller diameter of the outflowing arterioles compared to the bringing ones.
^ Stage 2 - reabsorption - Reabsorption of water and dissolved substances necessary for the body (amino acids, fine proteins, glucose, sodium, potassium, calcium, phosphates). Reabsorption occurs in the convoluted tubules of the first and second order. For a day, an adult forms 180 liters of primary urine, of which 178-179 liters are reabsorbed and only 1.0-1.5 liters of final urine is excreted. The second stage of urine formation ensures the concentration function of the kidneys, that is, the ability of the kidneys to concentrate primary urine.
^ Stage 3 - secretion in the urine by the epithelium of the convoluted tubules of hydrogen ions, potassium, ammonia, drugs, dyes. The secretion process helps to eliminate from the body all unnecessary substances formed as a result of metabolic processes, and ensures the final formation of urine.
^ Urine composition is normal. Urine is a liquid of complex chemical composition, in which about 150 substances are dissolved. Most of the urine (95%) is water, 5% is solid, of which 3.4% is organic and 1.6% is inorganic.
Urine organic matter is represented mainly by the end products of protein metabolism - urea, uric acid, creatinine. The urine also contains a small amount of enzymes, vitamins, pigments, hormones. About 40 g of organic matter is excreted in the urine per day. Urine inorganic substances include sodium, potassium, calcium, ammonia, etc.
^ Pathological urine impurities - components of urine, which are not normally contained in it, but appear only in case of diseases. Pathological impurities of urine include protein, glucose, acetone bodies, bilirubin, hemoglobin, etc. The presence of pathological impurities in the urine is indicated by special terms: proteinuria (protein in the urine), glucosuria (glucose in the urine), etc.
^ EXAMINATION OF URINE
General urine analysisis a widespread type of research that allows one to judge the nature and severity of the pathological process in the kidneys and urinary system.
The general analysis of urine includes three types of research.
1. Determination of the physical properties of urine: quantity, color, transparency, sediment, reaction, odor, relative density.
2. Chemical examination of urine:
Qualitative determination of protein and glucose, that is, determination of the presence of protein and glucose;
if protein and glucose are detected, their amount is determined.
A general urine test is performed in the morning, the most concentrated portion of urine.
The collection of urine is usually carried out by the patient himself after a thorough toilet of the external genitalia. A clean, wide-mouthed vessel with a lid is used to collect urine. Urine collected for general analysis can be stored in a cold place for no more than 1.5-2 hours.
In addition to the general analysis of urine, at the special request of the doctor, additional chemical studies of urine can be carried out to determine ketone bodies, urobilin, bilirubin, blood pigment - hemoglobin, etc., as well as quantitative methods of microscopic examination of urine sediment (according to Nechiporenko, Kakovsky-Addis, etc. .).
1.2.1. Study of the physical properties of urine
^ 1.2.1.1. AMOUNT OF URINE
A healthy adult has a daily amount of urine -daily urine output [from the Greek. diurēsisurination] is 0.8-1.5 liters.
The volume of the morning portion of urine (usually 150-250 ml) does not give an idea of the daily urine output. To determine daily urine output, it is necessary to examine daily urine (that is, urine collected within 24 hours).
In different conditions, the daily urine output may vary. An increase in daily urine output of more than 2 liters is called polyuria [from the Greek. polys many + urina urine] . It can be physiological (in healthy people in special conditions) and pathological (in diseases). Physiological polyuria is observed when drinking a lot of fluids and under stress. Pathological polyuria develops in chronic renal failure, pyelonephritis, resorption of edema. Severe polyuria (up to 3-4 years) is characteristic of diabetes mellitus. Especially sharp polyuria (up to 30 liters per day) is observed with diabetes insipidus (insufficiency of the pituitary antidiuretic hormone).
Oliguria [from the Greek. oligossmall amount +urina] - decrease in daily urine output less than 0.6l. It can also be physiological and pathological. Physiological oliguria occurs with restriction of drinking, loss of a large amount of fluid with sweat during significant physical exertion and high ambient temperatures. Pathological oliguria occurs with kidney disease (acute renal failure, acute glomerulonephritis), as well as with extrarenal fluid loss (vomiting, diarrhea, burn disease).
Anuria [from the Greek. a absence + urina] - complete cessation of urine excretion is true, which depends on the cessation of urine production by the kidneys (in acute renal failure), and mechanical - due to the presence in the urinary tract of a mechanical obstacle to the outflow of urine (stones, tumors).
Daily urine output is divided into daytime and nighttime. Normally, the ratio of daytime diuresis to nighttime is 3: 1 - 4: 1, that is, daytime diuresis is 3-4 times greater than nighttime. The predominance of nocturnal diuresis over daytime is called nocturia [from the Greek. nyx, nyktos night + urina] and is observed in chronic renal failure, prostate tumors.
Dysuria - painful urination [from the Greek.dys violation + urina] and pollakiuria – frequent urination [from the Greek.pollakis frequent + urina] are characteristic of cystitis (inflammation of the bladder).
COLOR OF URINE
Normal urine has a straw yellow color of varying intensity. The characteristic color of urine is given by the pigments contained in it:urochromes A and B, uroerythrin, stercobilinogen, which is commonly called in urine urobilin ... The intensity of the color of urine in healthy people depends on the amount of liquid drunk: with an increased drinking regime, the urine becomes lighter, and with restriction of drinking, increased sweating, it acquires a more intense yellow color. Some foods and medications can color the urine. Red (pink) color is given to urine by amidopyrine, aspirin, beets; brown - salol and naphthol; blue-green - methylene blue; brown - activated carbon, etc. The reasons for the change in the color of urine with pathology are presented in table 1.
Table 1
Causes of urine discoloration
| Urine color | Pathological condition | ^ Cause of color change |
| Dark yellow | Edema, vomiting, diarrhea, burn disease | High concentration of pigments |
| Pale, watery | Diabetes, diabetes insipidus | Low concentration of pigments |
| Red | Kidney stone disease (renal colic) | Hematuria (unaltered blood) |
| "Meat slop" | Acute glomerulonephritis cystitis | Hematuria (altered blood) |
| "Strong tea" | Hemolytic jaundice | Urobilinuria |
| "Beer" | Parenchymal jaundice | Bilirubinuria + urobilinuria |
| "Beer" | Obstructive jaundice | Bilirubinuria |
| Black | Hemolytic kidney | Hemoglobinuria |
| Whitish | Fatty degeneration of the kidneys | Drops of fat |
^ 1.2.1.3. TRANSPARENCY OF URINE
Normally, freshly excreted urine is clear. When standing, it becomes cloudy due to the precipitation of salts and cellular elements, the multiplication of bacteria.
table 2
Causes of cloudy urine and how to remove it
| ^ The reason for the turbidity of urine | Turbidity Removal Methods |
| Cellular elements: erythrocytes, leukocytes, epithelium | |
| Slime | Centrifugation, filtration |
| Fat | Add ether |
| Bacteria | Bacterial filter |
| Urata | Heating, adding alkali |
| Phosphates | Adding acetic acid |
| Oxalates | Adding hydrochloric acid |
In case of diseases, cloudy urine may be excreted. In these cases, turbidity can be caused by a large number of cellular elements (erythrocytes, leukocytes), bacteria, fat, and salts.
The clarity of urine is assessed by eye as: transparent, turbid, turbid.
^ Urine sedimentare formed during prolonged standing or when urine is cooled to 0˚С. Sediments can be composed of salts and cellular elements.
Macroscopically (that is, by eye) precipitation is described in three ways:
color (white, pink, brick red, etc.);
nature (amorphous, crystalline);
severity (abundant, insignificant).
^ 1.2.1. 4. REACTION OF URINE
Normal urine reaction is slightly acidic or neutral (pH = 5.0-7.0). In healthy people, the reaction of urine depends mainly on the food intake. From eating meat food, it shifts to the acidic side, and from plant products - to the alkaline one.
Table 3
The reasons for the change in urine reaction
^ Methods for determining the reaction of urine
Using indicator paper (universal indicator paper with a pH range of 1.0-10.0; special indicator paper for determining the pH of urine with a range of 5.0-8.0, combined test strips).
Standardized method with a liquid indicator bromothymol blue (pH determination range 6.0-7.6) according to Andreev.
Determination of the reaction of urine with the indicator bromothymol blue (according to Andreev)
Reagent: 0.1% solution of the indicator bromothymol blue.
^ Research progress. 1-2 drops of the indicator are added to 2-3 ml of urine. The color of the solution is used to judge the reaction of urine: yellow corresponds to an acidic reaction, brown to slightly acidic, herbaceous color to a neutral reaction, brown-green color to a slightly alkaline reaction, blue-green color to an alkaline reaction.
This test is very simple, but only gives a rough idea of the urine reaction. It is impossible to distinguish urine with normal pH from pathologically acidic by this method.
^ 1.2.1.5. URINE ODOR
It has no great diagnostic value. Normally, urine has a mild, specific odor.
With prolonged storage, accompanied by bacterial decomposition, urine acquires a pungent ammonia odor. The same smell has urine with cystitis. In diabetes mellitus, urine smells like acetone (rotten fruit) due to the presence of acetone bodies in it.
^ 1.2.1.6. RELATIVE DENSITY OF URINE
The relative density (specific gravity) of urine is proportional to the concentration of substances dissolved in it: urea, uric acid, creatinine, salts.
In healthy people, the relative density of urine fluctuates during the day from 1.005 to 1.030. In the morning, the most concentrated portion of urine, it is 1.020-1.026.
The relative density of urine is affected by the presence of pathological impurities in it - protein and glucose. Each 3 g / l of protein increases the relative density of urine by 1 division of the urometer (0.001), and every 10 g / l of glucose - by 4 divisions (0.004).
A low relative density of urine occurs in polyuria and chronic renal failure, and very high - up to 1.040-1.050 - most often in diabetes mellitus.
The relative density of urine gives an idea of the concentration ability of the kidneys, that is, the ability of the renal tubules to concentrate primary urine by reabsorbing water from it. The value of the relative density of the morning urine portion, equal to or greater than 1.018-1.020, indicates the preserved concentration function of the kidneys.
The relative density of urine is determined using a urometer - a special hydrometer with a scale from 1,000 to 1,050.
^ 1.2.1.7. ZIMNITSKY'S TEST
It is one of the methods for studying the functional state of the kidneys, it is used to assess the concentration of the kidneys. The test consists in dynamic monitoring of the amount and relative density of urine in 3-hour portions during the day. A prerequisite for conducting a test is a normal drinking regime, especially the exclusion of excessive fluid intake.
On the eve of the study, 8 cans are prepared. Mark them, indicating the name of the subject and the time of urine collection:
6-9 hours 5.18 p.m. to 9 p.m.
9-12 hours 6.21-24 hours.
12-15 hours. 7.0-3 hours.
15-18 hours 8.3-6 hours.
At 6 o'clock in the morning, the subject empties the bladder, but this portion of urine is not used for analysis. Then, every 3 hours during the day, the patient collects urine in jars with the appropriate time designation.
In the laboratory, in all 8 portions, the relative density and the exact amount of urine are determined using a graduated cylinder.
To evaluate the Zimnitsky sample, you must:
Calculate separately the daytime and nighttime urine output. Daytime diuresis is determined by summing up the amount of urine in the first 4 portions, and nighttime diuresis - in the last four;
Reveal the maximum and minimum relative density during the day and determine the difference between them (max ρ - min ρ).
The results of the Zimnitsky test are normal. Normal concentration function of the kidneys is characterized by: the ratio of daytime diuresis to nighttime 3: 1 - 4: 1; the difference between the maximum and minimum relative density is equal to or greater than 0.016.
A change in the concentration ability of the kidneys is indicated by a change in the ratio between day and night diuresis, nocturia, a decrease in the difference between the maximum and minimum relative density of urine, as well as isostenuria and hypostenuria.
Isostenuria [from the Greek. isos equal + urina] - excretion of urine during the day (in all 8 portions) with a constant relative density equal to the relative density of blood plasma - 1.010-1.011. Isosthenuria indicates a complete loss of concentration by the kidneys and is characteristic of chronic renal failure.
Hypostenuria [from the Greek. hypo below normal + urina] – excretion of urine during the day (in all 8 portions) with a constant relative density, less than the relative density of blood plasma, that is, less than 1.010. Hypostenuria indicates a sharp violation of the concentration function of the kidneys.
^ 1.2.1.8. CONTROL QUESTIONS ON THE TOPIC "RESEARCH OF THE PHYSICAL PROPERTIES OF URINE"
1. What tests are included in the general urinalysis?
2. How does the daily urine output change at high ambient temperatures?
3. What disease is characterized by pronounced polyuria?
4. What is hypostenuria?
5. What determines the value of the relative density of urine?
6. How is the relative density of urine determined?
7. What substances significantly increase the relative density of urine?
8. What is the true relative density of urine when the urometer reads 1.038 and contains 15 g / l of glucose in it?
9. What is the principle of Zimnitsky's test?
10. What stage of urine formation is characterized by Zimnitsky's test?
11. What is the characteristic of Zimnitsky's test in chronic renal failure?
12. What condition must be observed when carrying out the Zimnitsky test?
13. Name the pigments of normal urine.
14. What is the color of urine with bilirubinuria?
15. In what cases is the Zimnitsky test not carried out?
16. What are urates? How do they dissolve?
17. What values of urine pH are typical for diabetes mellitus?
18. What explains the alkaline reaction of urine in acute cystitis?
1.2.2. Chemical examination of urine
^ 1.2.2.1. DETERMINATION OF PROTEIN IN THE URINE
Normally, there is practically no protein in the urine. The presence of protein in the urine is calledproteinuria [from lat. protein protein + urina urine].
At the place of occurrence, renal (renal) proteinuria is distinguished, in which protein enters the urine from the kidneys, and extrarenal (extrarenal), when protein enters the urine from the urinary tract and genitals.
^ Renal proteinuria are divided into organic and functional.Organic renal proteinuria are observed in kidney diseases with damage to their structural unit - the nephron. Organic renal proteinuria is always persistent, long-term and is one of the main symptoms of the disease. They are found in acute and chronic glomerulonephritis, pyelonephritis, chronic renal failure, renal amyloidosis, nephrotic syndrome.
According to the mechanism of occurrence, organic renal proteinuria are glomerular and tubular. Glomerular proteinuria occurs due to increased permeability of the renal filter and can be massive (up to 10-20 g / l of protein). They are found in glomerulonephritis, renal amyloidosis, and toxic damage to the renal parenchyma. Depending on the ability of the renal filter to pass protein molecules of one size or another into the urine, glomerular proteinuria are divided into selective [from lat.selectioselection, selection] and non-selective. At selective proteinuria, only fine proteins with a relatively small molecular size (albumin) pass into the urine. With non-selective proteinuria, not only low molecular weight, but also high molecular weight proteins (globulins) pass into the urine, which indicates the severity of damage to the glomerular filter. The selectivity of proteinuria is judged by the results of the study of protein fractions of urine by electrophoresis.
Table 4
Causes and types of proteinuria
Tubular proteinuria develops with a decrease in protein reabsorption in the renal tubules (pyelonephritis). They usually do not exceed 2g / l.
Functional renal proteinuria are in healthy people under special circumstances:
Physical overexertion - "marching" proteinuria in soldiers after marches, sports proteinuria in athletes, etc.;
After severe hypothermia - cold;
After eating a large amount of raw egg white (alimentary) [from lat.alimentum nutrition];
In pregnant women in the last weeks before childbirth and in newborns in the first days of life.
All types of functional proteinuria do not last long. They quickly disappear when the circumstances that caused them disappear and usually do not exceed 1 g / l.
Conditionally, functional renal proteinuria also includes orthostatic and congestive proteinuria. Orthostatic proteinuria is otherwise called lordic [from lat.lordosforward curvature of the spine]. It is observed more often in adolescents of asthenic constitution with hyperlordosis of the lower segments of the thoracic spine. At the same time, the excretion of protein in the urine does not occur constantly, but only in the upright position of the body, hence the name - orthostatic [from lat.ortos straight + statusposition]. Orthostatic proteinuria develops as a result of the pressure of the curved spine on the vessels of the kidneys.
Congestive proteinuria occurs in patients with cardiovascular diseases, when, due to circulatory disorders, blood stagnation occurs in all internal organs, including the kidneys. The amount of protein in congestive proteinuria can reach 2-5 g / l.
^ Extrarenal proteinuria develop when protein enters the urine from the urinary tract and genitals - with inflammation of the bladder (cystitis), urethra (urethritis), vagina (colpitis). Extrarenal proteinuria depends on the admixture of secretions from the urogenital organs (leukocytes, erythrocytes).
^ Methods for determining protein in urine. Determination of protein is included in the general analysis of urine, being its mandatory component. First, a qualitative determination of the protein is carried out using:
Standardized sample with 20% sulfosalicylic acid solution;
Rapid tests of the "Albufan" type.
Normally, these samples are negative. If they give a positive result, that is, if protein is found in the urine, then its amount is determined. For the quantitative determination of protein in urine, standardized methods are used:
Turbidimetric with 3% sulfosalicylic acid solution;
Brandberg-Roberts-Stolnikov;
Biuret;
With pyrogallol red.
The amount of protein in the urine is expressed in g / l. Normally, the amount of protein in the urine does not exceed 0.033 g / l.
GOST R 53079.1-2008
Group P20
NATIONAL STANDARD OF THE RUSSIAN FEDERATION
Laboratory clinical technologies
QUALITY ASSURANCE OF CLINICAL LABORATORY STUDIES
Part 1
Rules for describing research methods
Medical laboratory technologies. Quality assurance of clinical laboratory tests.
Part 1. Rules for description of methods of clinical laboratory tests
OKS 11.020
Introduction date 2010-01-01
Foreword
The goals and principles of standardization in the Russian Federation are established by the Federal Law of December 27, 2002 N 184-FZ "On Technical Regulation", and the rules for the application of national standards of the Russian Federation - GOST R 1.0-2004 "Standardization in the Russian Federation. Basic Provisions"
Information about the standard
1 DEVELOPED by the Laboratory for the Problems of Clinical and Laboratory Diagnostics of the Moscow Medical Academy. I.M.Sechenov of Roszdrav, the Department of Clinical Laboratory Diagnostics and the Department of Biochemistry of the Russian Medical Academy of Postgraduate Education of the Russian Healthcare Agency, the Department of Certification and Quality Control of Clinical Laboratory Research of the State Scientific Center for Preventive Medicine of Rosmedtechnologies, the laboratory of biochemistry of amines and cyclic nucleotides of the Research Institute of Biomedical Chemistry of the Russian Academy medical sciences
2 INTRODUCED by the Technical Committee for Standardization TC 466 "Medical Technologies"
3 APPROVED AND PUT INTO EFFECT by Order of the Federal Agency for Technical Regulation and Metrology of the Russian Federation of December 18, 2008 N 464-st
4 INTRODUCED FOR THE FIRST TIME
Information about changes to this standard is published in the annually published information index "National standards", and the text of changes and amendments - in the monthly published information indexes "National standards". In case of revision (replacement) or cancellation of this standard, the corresponding notice will be published in the monthly published information index "National standards". Relevant information, notice and texts are also posted in the public information system - on the official website of the Federal Agency for Technical Regulation and Metrology on the Internet
1 area of use
1 area of use
This standard establishes the rules for describing in laboratory manuals, reference books and guidance materials for ready-made reagent kits (test systems) of clinical laboratory research methods intended for use in medical laboratories of all forms of ownership. This International Standard is intended for use by all organizations, institutions and enterprises, as well as individual entrepreneurs whose activities are related to the provision of medical care.
2 Normative references
This standard uses normative references to the following standards:
GOST R ISO 5725-2-2002 Accuracy (correctness and precision) of methods and results of measurements. Part 2. Basic method for determination of repeatability and reproducibility of a standard measurement method
GOST R ISO 9001-2008 Quality management systems. Requirements
GOST R ISO 15189-2006 Medical laboratories. Particular requirements for quality and competence
GOST R ISO 15193-2007 Medical devices for in vitro diagnostics. Measurement of quantities in samples of biological origin. Description of reference measurement procedures
GOST R ISO 15195-2006 Laboratory medicine. Requirements for reference measurement laboratories
GOST R ISO / IEC 17025-2006 General requirements for the competence of testing and calibration laboratories
GOST R ISO 17511-2006 Medical devices for in vitro diagnostics. Measurement of quantities in biological samples. Metrological traceability of values assigned to calibrators and control materials
GOST R ISO 18153-2006 Medical devices for in vitro diagnostics. Measurement of quantities in biological samples. Metrological traceability of enzyme catalytic concentration values assigned to calibrators and control materials
GOST R 53022.1-2008 Clinical laboratory technologies. Requirements for the quality of clinical laboratory research. Part 1. Quality management rules for clinical laboratory research
GOST R 53022.2-2008 Clinical laboratory technologies. Requirements for the quality of clinical laboratory research. Part 2. Assessment of analytical reliability of research methods (accuracy, sensitivity, specificity)
GOST R 53022.3-2008 Clinical laboratory technologies. Requirements for the quality of clinical laboratory research. Part 3. Rules for assessing the clinical information content of laboratory tests
GOST R 53022.4-2008 Clinical laboratory technologies. Requirements for the quality of clinical laboratory research. Part 4. Rules for the development of requirements for the timeliness of the provision of laboratory information
GOST 7601-78 Physical optics. Terms, letter designations and definitions of basic quantities
Note - When using this standard, it is advisable to check the validity of reference standards in the public information system - on the official website of the Federal Agency for Technical Regulation and Metrology on the Internet or according to the annually published index "National Standards", which was published as of January 1 of the current year, and according to the relevant monthly information signs published in the current year. If the reference standard is replaced (changed), then when using this standard, the replacement (modified) standard should be followed. If the reference standard is canceled without replacement, then the provision in which the reference to it is given applies to the extent not affecting this reference.
3 Rules for describing research methods and test systems intended for use in medical laboratories
3.1 General
Modern analytical capabilities of laboratory medicine are represented by a wide variety of research methods that can be used to detect and / or measure the same analyte, biological object. However, the actual values of the results of these studies performed by different methods can differ significantly from each other, which can lead to incomparability of the results of the patient examination performed in different institutions, and their erroneous interpretation, in particular, when a patient is transferred from one medical institution to another. An accurate characterization of the properties of the research method, based on uniform standardized data on the details of analytical procedures, the properties of the analysis tools used, the characteristics of the analytical reliability and clinical information content of the study, should be used when choosing and reproducing a method in clinical diagnostic laboratories, to facilitate an objective comparison of the results of using various methods and prevention of errors in the interpretation of research carried out in the laboratories of various medical organizations.
3.2 Analytical properties of research methods
The analytical properties of the method used for the study of biological material are of decisive importance for the quality of the study. According to the national standards GOST R ISO 9001, GOST R ISO 15189 and GOST R ISO / IEC 17025, in a medical laboratory, the quality must be ensured by analytical procedures, including the properties of the methods used.
According to the characteristics and form of expression of the result obtained (GOST R ISO 15193), the methods of clinical laboratory research are subdivided:
- to quantitative ones that measure quantities, giving results in a scale of differences or a scale of relations, where each value is a numerical value multiplied by a unit of measurement (in a series of values, ordinary statistical parameters can be calculated: arithmetic mean, standard deviation, geometric mean and coefficient of variation );
- semi-quantitative, the results of which are expressed in an ordinal (ordinal) scale, in which the values can be expressed by phrases or numbers expressing the size of the corresponding properties, and used for ranking, however, the differences and ratios on the scale do not matter for comparison [for a number of values may be calculated fracttiles (including the median) and applied some nonparametric tests, for example, the Kolmogorov-Smirnov, Wilcoxon tests and the sign test].
Ensure the conduct of studies of samples of biomaterials of patients in accordance with the needs of the clinic for information content, analytical reliability and timely receipt of research results, established by the relevant regulatory documents of the quality management system for clinical laboratory research (GOST R 53022.4);
- to ensure the comparability of the results of studies of analytes and biological objects carried out in various health care organizations, that is, to be standardized with respect to the description and characteristics of their analytical principles and implemented technologies;
- be economically acceptable for medical organizations.
When describing research methods and test systems intended for use in clinical diagnostic laboratories of medical organizations, reliable data borrowed from special scientific literature obtained in accredited expert laboratories, or developers' own data regarding:
- metrological traceability of the analytical properties of the proposed methods to the properties of the reference research methods in accordance with GOST R ISO 15193 and GOST R ISO 17511 (if international reference methods are available);
- characteristics of the properties of the analysis tools used;
- evaluating the efficiency of the practical application of the method.
3.3 Scheme for a standardized description of a working method for clinical laboratory research
3.3.1 General
This International Standard provides a general framework for a standardized description of a test method. Descriptions of the procedures for the methods of research of individual analytes used in the provision of the corresponding simple or complex medical services are given in the regulatory documents for the technologies of specific medical laboratory services.
A standardized description of a clinical laboratory research method is a set of clear and complete descriptions of interrelated analytical procedures of a physical, chemical, biological nature; conditions for their implementation; reagents and equipment, the use of which, in accordance with their description, ensures reliable detection / determination of the desired analyte or biological object in a sample of biological material.
3.3.2 Scheme of a standardized method description
The standardized description of the method should contain the following data:
a) the name of the method indicating the desired analyte, biological object;
b) the principle of detection or determination of an analyte, a biological object in this method;
c) the necessary chemical, biological reagents and characteristics of their physicochemical, biological properties (in the case of using separate reagents):
1) degree of purity (qualification) - for chemical reagents;
2) the range of activity - for enzymes, specificity - for enzyme substrates according to GOST R ISO 18153; specificity and affinity for antibodies;
3) the composition of the components - for nutrient media;
4) detection wavelength range - for chromophores, fluorophores;
5) the composition and characteristics of the components, ionic strength, pH - for buffer solutions.
When using ready-made forms of reagent kits, the principle of the method, the composition of the reagents, the presence of state registration, compliance with the requirements of analytical reliability, metrological traceability and commutability of the calibrator, and the method of application are indicated. For all reagents - the period of stability in dry form and after dissolution, peculiarities of storage conditions, degree of toxicity and biological hazard.
3.3.3 Special equipment for sample preparation and analysis
Equipment for sample preparation and analysis:
- manual,
- semi-automatic,
- automatic.
Characteristics of instruments and equipment required to ensure the implementation of the study:
- for dispensers - the required volume and dosing accuracy;
- for centrifuges - the appropriate operating mode (revolutions per minute, rotor rotation radius, the need for cooling);
- for thermostats - the temperature during operation and the permissible limits of its fluctuations;
- for sterilization equipment - pressure and temperature during operation, the limits of their fluctuations;
- for anaerostats - CO content;
- for optical measuring instruments - type of photometry: absorption, flame, horizontal, vertical, reflective, turbidimetry, nephelometry, fluorometry, luminometry, time-resolved fluorometry - the corresponding wavelength, slit width, light transmission, thickness of the absorbing layer of the colored solution (internal cuvette size, cm) by; when using a thermostated cuvette - the set temperature and the permissible limits of its fluctuations);
- for microscopes - type of microscopy, magnification, resolution in accordance with GOST R 7601,;
- for devices for electrophoresis - the composition of the buffer solution, voltage and current strength, type of carrier;
- for devices for chromatography - composition and characteristics of stationary and mobile phases, type of detector;
- for devices based on the electrochemical principle of measurement, - signal parameters, type of detector;
- for coagulometers - the principle of operation, the method of detection;
- for flow cytometers - the principle of operation, measured and calculated parameters;
- systems for image analysis should be characterized by a database, the main criteria for evaluating images.
For all instruments that are measuring instruments, their metrological characteristics must be given.
3.3.4 Analyte testing
When describing the study of the analyte, indicate:
a) investigated (analyzed) biological material: biological fluid, excreta, tissue;
b) specific pre-analytical precautions at the pre-laboratory and intra-laboratory stages:
1) a sample of the material under study: place, method, conditions, time of taking, volume;
2) the material of the containers for taking samples, depending on the properties of the desired analyte, the procedure for processing the biomaterial;
3) additives: anticoagulants, preservatives, fixatives, gels; the amount of additives in relation to the volume of the sample;
4) storage and transportation conditions, taking into account the stability characteristics of the analyte: light, temperature, sterility, isolation from the ambient atmosphere, maximum storage duration;
5) description of the sample preparation procedure;
c) analysis progress:
1) procedures and their conditions: temperature of the reaction, pH, time intervals for individual stages of the analysis procedures (incubation, the duration of the delay in the reaction to the linear section, the duration of the linear section of the reaction), the type of blank sample (matrix, reagents, mixing sequence); measured material: sample (biomaterial plus reagents); sample volume required for this measurement option, the ratio of biomaterial and reagents by volume, stability of the reaction product;
2) calibration (calibration) procedures: calibration material, traceability of its properties to the properties of a certified standard sample (international certified reference material); plotting and characterizing a calibration graph, linearity area, calibration factor, analyte detection limit, measurement range; nonlinear calibration plots; methods of calculating the results;
d) assessment of the analytical reliability of the method: correctness, precision (repeatability and reproducibility), analytical sensitivity, analytical specificity; Recommended materials for assessing the accuracy and precision of an analytical method; comparison with the requirements for the analytical quality of the determination of a given analyte; possible sources of errors of various types, measures for their elimination.
If there is a reference method, an assessment in relation to this method in accordance with GOST R ISO 15193. Possible interferences: drugs, hemolysis, icterus samples, lipemia;
e) assessment or calculation of the research result:
1) mathematical rules for calculating the result; presentation of the result: in units of the International System of Units and in traditionally used units (for quantitative methods); for semi-quantitative - in the ordinal (ordinal) scale; for non-quantitative ones - in the form adopted for this type of research (positive or negative result; the desired analyte was found or not; in a descriptive (nominal) form - for cytological studies);
2) the reference interval, including sex and age characteristics; analyte personality index (to assess the applicability of comparison with a reference interval); forms of pathology, for the diagnosis of which the method of research of a given analyte, biological object is intended;
3) technical and economic assessment, taking into account the consumption of materials, costs of working time, depreciation of equipment (if possible, per unit of clinical information obtained during the study);
4) source of data on the characteristics of the method: the organization that carried out the assessment; expert laboratory; the result of an interlaboratory (multicenter) experiment to evaluate the method; regulatory document of a competent national or international organization.
3.4 Requirements for the description of the standardized method
Manufacturers should comply with certain requirements when describing analytical tools (reagent kits and instruments) for a standardized analyte test method.
3.4.1 The scheme of a standardized description of a research method should be detailed, since it is designed to describe the methods of various types of research used in clinical diagnostic laboratories of medical organizations.
When describing a specific method, those positions should be reflected that are necessary to characterize the analytical procedures and analytical tools inherent in the study of this type.
Note - The right to omit some characteristics of reagents in their ready-made kits, due to the protection of intellectual property, does not apply to data on the critical parameters of the method: sensitivity, specificity, correctness, metrological traceability, precision, linearity, measurement interval.
3.4.2 When describing a research method based on the use of analytical tools (reagent kits, instruments) manufactured by a specific manufacturing organization and being a closed system, the characteristics of the accuracy and precision of the results obtained in comparison with the reference research method or the method chosen for comparison should be given. the properties of which are compared with the reference method, data on the commutability of the calibrator.
3.4.3 With regard to measuring instruments proposed for use when performing this research method, the federal executive body in the field of technical regulation and metrology * carries out state metrological control and supervision.
________________
* Federal Law of June 26, 2008 N 102-FZ "On ensuring the uniformity of measurements".
State metrological control includes:
- type approval of measuring instruments;
- verification of measuring instruments, including standards;
- licensing of the activities of legal entities and individuals for the manufacture and repair of measuring instruments.
State metrological supervision is carried out:
For the release, condition and use of measuring instruments;
- certified measurement procedures;
- standards of units of quantities;
- compliance with metrological rules and norms *.
________________
* The functions of state metrological control and supervision are carried out by the Federal Agency for Technical Regulation and Metrology.
In the description of the standardized method for clinical laboratory research, information on registration with the authorized state body and entry into the state register, for measuring devices - on registration with the national technical regulation body, if there is a technical regulation for devices of this type - on the mark must be provided. compliance.
3.4.4 Ready-made reagent kits for this research method must be tested in accordance with the established procedure, meet the relevant technical requirements and must be entered in the state register, information on registration and permission for use must be provided in the description of the analyte research method.
Bibliography
ISO 8036: 1998 Optics and optical instruments - Microscopes |
|
ISO 8039: 1997 Optics and optical instruments - Magnification microscopes |
|
World Health Organization. Anticoagulant use and stability of blood, serum and plasma samples. - Geneva, 2002 |
Electronic text of the document
prepared by Kodeks CJSC and verified by:
official publication
M .: Standartinform, 2009
A large number of existing diseases, the individual degree of different people complicate the diagnostic process. Often, in practice, it is not enough to use only the knowledge and skills of a doctor. In this case, clinical laboratory diagnostics helps to make the correct diagnosis. With its help, pathologies are detected at an early stage, the development of the disease is monitored, its possible course is assessed and the effectiveness of the prescribed treatment is determined. Today, medical laboratory diagnostics is one of the most rapidly developing areas of medicine.
Concept
Laboratory diagnostics is a medical discipline that applies in practice standard methods for detecting and monitoring diseases, as well as looking for and studying new methods.
Clinical laboratory diagnostics greatly facilitates and allows you to choose the most effective therapy regimen.
Sub-branches of laboratory diagnostics are:
The information obtained using various methods of clinical laboratory diagnostics reflects the course of the disease at the organ, cellular and molecular levels. Due to this, the doctor has the opportunity to diagnose pathology in a timely manner or evaluate the result after the treatment.
Tasks
Laboratory diagnostics is designed to solve the following tasks:
- continuous search and study of new methods of analysis of biomaterial;
- analysis of the functioning of all human organs and systems using existing methods;
- detection of a pathological process at all its stages;
- control over the development of pathology;
- evaluation of the result of therapy;
- precise definition of the diagnosis.
The main function of the clinical laboratory is to provide the doctor with information about the analysis of biomaterial, comparing the results with normal values.
Today, 80% of all information important for diagnosis and treatment monitoring is provided by the clinical laboratory.
Types of test material
Laboratory diagnostics is a way to obtain reliable information by examining one or several types of human biological material:
- Venous blood is taken from a large vein (mainly at the bend of the elbow).
- Arterial blood - most often taken to assess CBS from large veins (mainly from the thigh or the area under the collarbone).
- Capillary blood is taken from a finger for a variety of studies.
- Plasma - it is obtained by centrifuging blood (i.e., separating it into its components).
- Serum - blood plasma after separation of fibrinogen (a component that is an indicator of blood clotting).
- Morning urine - collected immediately after waking up, is intended for general analysis.
- Daily urine output is urine that is collected in one container during the day.
Stages
Laboratory diagnostics includes the following steps:
- preanalytic;
- analytical;
- post-analytical.
The preanalytical stage implies:
- Compliance by a person with the necessary rules for preparing for analysis.
- Documentary registration of the patient upon arrival at the medical institution.
- Signature of tubes and other containers (for example, with urine) in the presence of the patient. The name and type of analysis are applied on them by the hand of a medical worker - he must pronounce these data aloud to confirm their reliability by the patient.
- Subsequent processing of the taken biomaterial.
- Storage.
- Transportation.
The analytical stage is the process of direct examination of the obtained biological material in the laboratory.
The post-analytical stage includes:
- Documentary registration of results.
- Interpretation of results.
- Formation of a report containing: data of the patient, the person who conducted the study, the medical institution, the laboratory, the date and time of the sampling of the biomaterial, the normal clinical limits, the results with the corresponding conclusions and comments.
Methods
The main methods of laboratory diagnostics are physical and chemical. Their essence lies in the study of the material taken for the relationship of its various properties.
Physicochemical methods are subdivided into:
- optical;
- electrochemical;
- chromatographic;
- kinetic.
The optical method is most commonly used in clinical practice. It consists in fixing changes in the light beam passing through the biomaterial prepared for research.
In second place in terms of the number of analyzes carried out is the chromatographic method.
Probability of errors
It is important to understand that clinical laboratory diagnostics is a type of research in which mistakes can be made.
Each laboratory must be equipped with quality instruments, analyzes must be performed by highly qualified specialists.
According to statistics, the main share of errors occurs at the preanalytical stage - 50-75%, at the analytical stage - 13-23%, at the post-analytical stage - 9-30%. Regular measures should be taken to reduce the likelihood of errors at each stage of laboratory research.
Clinical laboratory diagnostics is one of the most informative and reliable ways to obtain information about the health of the body. With its help, it is possible to identify any pathologies at an early stage and take timely measures to eliminate them.
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The methods of biochemical, coagulological, serological, immunological, morphological, mycological, cytological studies of human body fluids are described. The book provides modern information about the structure and function of vital organs, clinical and laboratory tests reflecting the features of their condition, methods of laboratory diagnostic research, about the peculiarities of changes in the biochemical and morphological composition of blood, urine, gastric contents, cerebrospinal fluid, sputum, genital discharge and other biological material for widespread diseases, as well as on the performance of quality control of laboratory tests, interpretation of the results. The description of each method includes information about the principle, the course of the study and the clinical and diagnostic value of the test performed. The book can be successfully used in teaching and practical activities of specialists in clinical laboratory diagnostics with secondary and higher medical education. Publisher: "MEDpress-inform" (2016) Format: 216.00mm x 145.00mm x 38.00mm, 736 pages
ISBN: 978-5-00030-273-6 |
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