Qualitative determination of protein in urine. Urine protein (proteinuria). Determination of protein in urine

21.06.2019

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A healthy person excretes 1.0–1.5 liters of urine per day. The content of 8–10 mg/dl of protein in it is a physiological phenomenon. The daily rate of protein in the urine of 100-150 mg should not arouse suspicion. Globulin, mucoprotein and albumin are what make up the total protein in the urine. A large outflow of albumin indicates a violation of the filtration process in the kidneys and is called proteinuria or albuminuria.

Each substance in the urine is assigned a “healthy” norm, and if the protein index fluctuates, this may indicate kidney pathology.

A general urinalysis implies the use of either the first (morning) portion, or a daily sample is taken. The latter is preferable for assessing the level of proteinuria, since the protein content has pronounced daily fluctuations. Urine during the day is collected in one container, the total volume is measured. For a laboratory that analyzes urine for protein, a standard sample (50 to 100 ml) from this container is sufficient, the rest is not required. For more information, an additional Zimnitsky test is carried out, which shows whether urine indicators per day are normal.

Methods for determining protein in urine
View	Subspecies			Peculiarities
quality	Geller test			The study of urine for the presence of protein
	Sulfosalicylic acid test
	Boil Analysis
quantitative		Turbidimetric	Protein from the urine interacts with the reagent, resulting in a decrease in its solubility. As reagents, sulfosalicylic and trichloroacetic acids, benzethonium chloride are used.
		Colorimetric	With some substances, the protein from the urine changes color. This is the basis of the biuret reaction and the Lowry method. Other reagents are also used - brilliant blue, pyrogallol red.
semi-quantitative			Give a relative idea of the amount of protein, the result is interpreted by the color change of the sample. Semi-quantitative methods include test strips and the Brandberg-Roberts-Stolnikov method.

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Protein in the urine is normal in an adult should not exceed 0.033 g / l. In this case, the daily rate is not higher than 0.05 g / l. For pregnant women, the protein norm in daily urine is more - 0.3 g / l., and in the morning urine the same - 0.033 g / l. The norms of protein in the general analysis of urine and in children differ: 0.036 g / l for the morning portion and 0.06 g / l per day. Most often, in laboratories, analysis is carried out by two methods, which show how much the protein fraction is in urine. The above norm values are valid for the analysis carried out with sulfosalicylic acid. If pyrogallol red dye was used, the values will differ by a factor of three.

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filtration in the renal glomeruli goes the wrong way;
absorption in the tubules of the protein is impaired;
some diseases put a heavy burden on the kidneys - when the protein in the blood is elevated, the kidneys simply “do not have time” to filter it.

The remaining causes are considered non-renal. This is how functional albuminuria develops. Protein in the urine test appears with allergic reactions, epilepsy, heart failure, leukemia, poisoning, myeloma, chemotherapy, and systemic diseases. Most often, it is this indicator in the patient's analyzes that will be the first bell of hypertension.

An increase in protein in urine may be due to non-pathological factors, so additional tests will be required.

Quantitative methods for determining protein in urine give errors, so it is recommended to conduct several analyzes, and then use the formula to calculate the correct value. The protein content in the urine is measured in g/l or mg/l. These protein values make it possible to determine the level of proteinuria, suggest a cause, assess the prognosis, and decide on a strategy.

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For the full functioning of the body, a constant exchange between blood and tissues is necessary. It is possible only if there is a certain osmotic pressure in the blood vessels. Blood plasma proteins just maintain such a level of pressure when low-molecular substances easily pass from an environment with a high concentration of them to an environment with a lower one. The loss of protein molecules leads to the release of blood from its channel into the tissues, which is fraught with severe edema. This is how moderate and severe proteinuria manifests itself.

The initial stages of albuminuria are asymptomatic. The patient pays attention only to the manifestations of the underlying disease, which is the cause of protein in the urine.

Trace proteinuria is called an increase in the level of protein in urine due to the use of certain foods.

Urine for analysis is collected in a clean, fat-free container. Before collection, the toilet of the perineum is shown, you need to wash it with soap. Women are advised to cover the vagina with a piece of cotton wool or a tampon so that vaginal discharge does not affect the result. On the eve it is better not to drink alcohol, mineral water, coffee, spicy, salty and food that gives urine color (blueberries, beets). Strong physical exertion, prolonged walking, stress, fever and sweating, excessive consumption of protein foods or drugs before passing urine provoke the appearance of protein in the urine of a completely healthy person. This tolerable phenomenon is called trace proteinuria.

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Kidney disease leading to protein loss:

Amyloidosis. Normal cells in the kidneys are replaced by amyloids (protein-saccharide complex), which prevents the organ from working normally. At the proteinuric stage, amyloids are deposited in the renal tissues, destroying the nephron and, as a result, the renal filter. This is how protein passes from the blood into the urine. This stage can last more than 10 years.
diabetic nephropathy. Due to improper metabolism of carbohydrates and lipids, the destruction of blood vessels, glomeruli and tubules in the kidney occurs. Protein in the urine is the first sign of a predictable complication of diabetes.
Diseases of inflammatory origin - nephritis. Most often, the lesions affect the blood vessels, glomeruli and pelvicalyceal systems, disrupting the normal course of the filtration system.
Glomerulonephritis in most cases is autoimmune in nature. The patient complains of a decrease in the amount of urine, back pain and increased pressure. For the treatment of glomerulonephritis, diet, regimen and drug therapy are recommended.
Pyelonephritis. In the acute period, it proceeds with symptoms of a bacterial infection: chills, nausea, headache. This is an infectious disease.
Polycystic kidney disease.

In a healthy body, protein molecules (and they are quite large in size) are not able to pass through the filtration system of the kidneys. Therefore, there should be no protein in the urine. This figure is the same for both men and women. If the analysis indicates proteinuria, it is important to consult a doctor to find out the reasons. The specialist will assess how elevated the protein level is, whether there is a concomitant pathology, how to restore the normal functioning of the body. According to statistics, a woman has a higher risk of developing a urinary tract infection than a man.

Method principle based on protein coagulation in urine in the presence of nitric (or 20% sulfosalicylic acid) acid.

Progress: 1-2 drops of nitric (or sulfosalicylic) acid are added to 5 drops of urine. If there is protein in the urine, turbidity appears.

Table. Detection of pathological components of urine .

Note: in the presence of glucose and protein in the test urine, their quantitative content is determined.

Method principle : when the protein interacts with pyrogallol red and sodium molybdate, a colored complex is formed, the color intensity of which is proportional to the protein concentration in the sample.

Reagents: Working reagent - pyrogallol red solution in succinate buffer, protein calibration solution with a concentration of 0.50 g/l

Progress:

Mix samples, hold for 10 minutes. at room temperature (18 -25ºС). Measure the optical density of the experimental (Dop) and calibration sample (Dk) against the control sample at λ=598 (578-610) nm. The color is stable for 1 hour.

Calculation: protein concentration in urine (С) g/l to calculate by the formula:

С= Dop/Dk×0.50

where: Dop \u003d Dk \u003d C \u003d g / l.

Normal values: up to 0.094 g/l, (0.141 g/day)

Conclusion:

Method principle : When D-glucose is oxidized by atmospheric oxygen under the action of glucose oxidase, an equimolar amount of hydrogen peroxide is formed. Under the action of peroxidase, hydrogen peroxide oxidizes chromogenic substrates (a mixture of phenol and 4 aminoantipyrine - 4AAP) with the formation of a colored product. The color intensity is proportional to the glucose content.

glucose oxidase

Glucose + O2 + H2O gluconolactone + H2O2

peroxidase

2H2O2 + phenol + 4AAP colored compound + 4H2O

Progress: add 1 ml of working solution and 0.5 ml of phosphate buffer into two test tubes. 0.02 ml of urine is added to the first tube, 0.02 ml of the calibrator (calibration, standard glucose solution, 10 mmol/l) is added to the second tube. The samples are mixed, kept for 15 minutes at a temperature of 370C in a thermostat, and the optical density of the experimental (Dop) and calibration (Dc) samples is measured against the working reagent at a wavelength of 500-546 nm.

Calculation: С = Dop/Dk  10 mmol/l Dop= Dk =

Conclusion:

Note. If the sugar content in the urine is more than 1%, it must be diluted.

Currently, biochemical laboratories use a unified express method for testing urine for glucose using reactive paper for glucose Glucotest or using combined test strips for pH, protein, glucose, ketone bodies and blood. Test strips are lowered into a vessel with urine for 1 second. and compare the color scale.

Protein determination using pyrogallol red indicator

The principle of the method is based on the photometric measurement of the optical density of a solution of a colored complex formed by the interaction of protein molecules with molecules of the Pyrogallol Red-Molybdate complex dye complex in an acidic medium. The color intensity of the solution is proportional to the protein content in the test material. The presence of detergents in the reagent provides an equivalent determination of proteins of different nature and structure.

Reagents. 1) 1.5 mmol/l solution of pyrogallol red (PGD): 60 mg of PGA are dissolved in 100 ml of methanol. Store at a temperature of 0–5 °C; 2) 50 mmol/l succinate buffer pH 2.5: 5.9 g succinic acid (HOOC-CH2-CH2-COOH); 0.14 g of sodium oxalate (Na2C2O4) and 0.5 g of sodium benzoate (C6H5COONa) are dissolved in 900 ml of distilled water; 3) 10 mmol/l solution of sodium molybdate crystal hydrate (Na2MoO4 × 2H2O): 240 mg of sodium molybdate are dissolved in 100 ml of distilled water; 4) Working reagent: 40 ml of PGA solution and 4 ml of sodium molybdate solution are added to 900 ml of succinate buffer solution. The pH of the solution is adjusted to 2.5 with 0.1 mol/l hydrochloric acid (HCl) solution and the volume is adjusted to 1 liter. The reagent in this form is ready for use and is stable when stored in a place protected from light and at a temperature of 2–25 °C for 6 months; 5) 0.5 g/l albumin standard solution.

Definition progress. 0.05 ml of the test urine is added to the first tube, 0.05 ml of the albumin standard solution is added to the second tube, and 0.05 ml of distilled water is added to the third tube (control sample), then 3 ml of the working reagent are added to these tubes. The contents of the tubes are mixed and after 10 minutes the sample and the standard are photometered against the control sample at a wavelength of 596 nm in a cuvette with an optical path length of 10 mm.

Calculation of the protein concentration in the test urine sample is carried out according to the formula:

where C is the concentration of protein in the test urine sample, g/l; Apr and Ast - extinction of the studied urine sample and albumin standard solution, g/l; 0.5 - concentration of standard solution of albumin, g/l.

Notes:

the color of the solution (color complex) is stable for one hour;
the directly proportional relationship between the protein concentration in the test sample and the absorbance of the solution depends on the type of photometer;
when the protein content in the urine is above 3 g/l, the sample is diluted with isotonic sodium chloride solution (9 g/l) and the determination is repeated. The degree of dilution is taken into account when determining the protein concentration.

See also:

Determination of protein in urine
Standardized test with sulfosalicylic acid
Unified Brandberg–Roberts–Stolnikov method
Determination of the amount of protein in the urine by reaction with sulfosalicylic acid
Biuret method
Detection of Bence-Jones protein in urine

Proteinuria is a phenomenon in which protein is determined in the urine, which indicates the possibility of damage to the kidneys, serves as a factor in the development of diseases of the heart, blood, and lymphatic vessels.

The detection of protein in urine does not always indicate a disease. A similar phenomenon is typical even for absolutely healthy people, in whose urine protein can be determined. Hypothermia, physical activity, the use of protein foods leads to the appearance of protein in the urine, which disappears without any treatment.

During screening, protein is detected in 17% of apparently healthy people, but only 2% of this number of people a positive test result is a sign of kidney disease.

Protein molecules should not enter the blood. They are vital for the body - they are the building material for cells, participate in reactions as coenzymes, hormones, antibodies. In both men and women, the norm is the complete absence of protein in the urine.

The function of preventing the body from losing protein molecules is performed by the kidneys.

There are two systems of the kidneys that filter urine:

renal glomeruli - do not let large molecules through, but do not retain albumins, globulins - a small fraction of protein molecules;
renal tubules - adsorb proteins filtered by the glomeruli, return back to the circulatory system.

Albumins (about 49%), mucoproteins, globulins are found in the urine, of which immunoglobulins account for about 20%.

Globulins are high molecular weight whey proteins that are produced by the immune system and the liver. Most of them are synthesized by the immune system, refer to immunoglobulins or antibodies.

Albumins are the fraction of proteins that are the first to appear in the urine already with minor kidney damage. A certain amount of albumin is also in healthy urine, but it is so insignificant that it is not detected using laboratory diagnostics.

The lower threshold, which can be detected using laboratory diagnostics, is 0.033 g / l. If more than 150 mg of protein is lost per day, then they speak of proteinuria.

Key facts about protein in urine

The disease with a mild degree of proteinuria is asymptomatic. Visually, urine that does not contain protein cannot be distinguished from urine, which contains a small amount of protein. Somewhat foamy urine becomes already with a high degree of proteinuria.

It is possible to assume active excretion of protein in the urine by the appearance of the patient only with a moderate or severe degree of the disease by the appearance of edema of the extremities, face, and abdomen.

In the early stages of the disease, indirect signs of proteinuria can be symptoms:

changes in the color of urine;
increasing weakness;
lack of appetite;
nausea, vomiting;
bone pain;
drowsiness, dizziness;
elevated temperature.

The appearance of such signs should not be ignored, especially during pregnancy. This may mean a slight deviation from the norm, and may be a symptom of developing preeclampsia, preeclampsia.

Quantifying protein loss is not an easy task; several laboratory tests are used to achieve a more complete picture of the patient's condition.

Difficulties in choosing a method for detecting excess protein in the urine are explained by:

low protein concentration, for the recognition of which high-precision instruments are required;
the composition of urine, which complicates the task, since it contains substances that distort the result.

The greatest information can be obtained from the analysis of the first morning portion of urine, which is collected after waking up.

On the eve of the analysis, the following conditions must be observed:

do not use spicy, fried, protein foods, alcohol;
avoid taking diuretics for 48 hours;
limit physical activity;
carefully follow the rules of personal hygiene.

Morning urine is the most informative, since it stays in the bladder for a long time, and to a lesser extent depends on food intake.

It is possible to analyze the amount of protein in the urine by a random portion, which is taken at any time, but such an analysis is less informative, and the probability of error is higher.

To quantify the daily loss of protein, an analysis of the total daily urine is done. To do this, within 24 hours, all the urine excreted during the day is collected in a special plastic container. You can start collecting at any time. The main condition is exactly one day of collection.

The qualitative definition of proteinuria is based on the property of the protein to denature under the influence of physical or chemical factors. Qualitative methods are screening methods that allow you to determine the presence of protein in the urine, but do not make it possible to accurately assess the degree of proteinuria.

Samples used:

with boiling;
sulfosalicylic acid;
nitric acid, Larionova's reagent with the Heller ring test.

A test with sulfosalicylic acid is performed by comparing a control urine sample with an experimental one, in which 7-8 drops of 20% sulfosalicylic acid are added to urine. The conclusion about the presence of the protein is made by the intensity of the opalescent turbidity that appears in the test tube during the reaction.

More often, the Geller test is used using 50% nitric acid. The sensitivity of the method is 0.033 g/l. At this concentration of protein in a test tube with a urine sample and a reagent, a white filamentous ring appears 2-3 minutes after the start of the experiment, the formation of which indicates the presence of the protein.

Geller test

Semi-quantitative methods include:

method for determining protein in urine with test strips;
Brandberg-Roberts-Stolnikov method.

The Brandberg-Roberts-Stolnikov determination method is based on the Geller ring method, but allows a more accurate assessment of the amount of protein. When performing a test according to this method, several dilutions of urine achieve the appearance of a filamentous protein ring in the time interval between 2-3 minutes from the start of testing.

In practice, the method of test strips with applied dye bromophenol blue as an indicator is used. The disadvantage of test strips is the selective sensitivity to albumin, which leads to a distortion of the result in the event of an increase in the concentration of globulins or other proteins in the urine.

The disadvantages of the method also include the relatively low sensitivity of the test to protein. The test strips begin to react to the presence of protein in the urine at a protein concentration exceeding 0.15 g / l.

Quantification methods can be conditionally divided into:

turbidimetric;
colorimetric.

The methods are based on the property of proteins to reduce solubility under the action of a binding agent with the formation of a poorly soluble compound.

Protein binding agents can be:

sulfosalicylic acid;
trichloroacetic acid;
benzethonium chloride.

The results of the tests are concluded based on the degree of attenuation of the light flux in the suspension sample compared to the control. The results of this method can not always be attributed to reliable due to differences in the conditions of carrying out: the speed of mixing the reagents, temperature, acidity of the medium.

Influence the assessment of taking medications the day before, before conducting tests according to these methods, you can not take:

antibiotics;
sulfonamides;
iodine preparations.

The method is affordable, which allows it to be widely used for screening. But more accurate results can be obtained using more expensive colorimetric techniques.

Colorimetric techniques are among the sensitive methods for accurately determining the concentration of protein in the urine.

To do this with high accuracy allow:

biuret reaction;
Lowry's technique;
staining techniques that use dyes that form complexes with urine proteins that differ visually from the sample.

Colorimetric methods for detecting protein in urine

The method is reliable, highly sensitive, allowing the determination of albumin, globulins, paraproteins in urine. It is used as the main way to clarify controversial test results, as well as daily protein in urine in patients with nephrology departments of hospitals.

Even more accurate results can be achieved by the Lowry method, which is based on the biuret reaction, as well as the Folin reaction, which recognizes tryptophan and tyrosine in protein molecules.

To eliminate possible errors, the urine sample is purified by dialysis from amino acids, uric acid. Errors are possible with the use of salicylates, tetracyclines, chlorpromazine.

The most accurate way to determine a protein is based on its ability to bind to dyes, of which are used:

ponceau;
coumasi brilliant blue;
pyrogall red.

During the day, the amount of protein excreted in the urine changes. In order to more objectively assess the loss of protein in the urine, the concept of daily protein in the urine is introduced. This value is measured in g/day.

For a quick assessment of the daily protein in the urine, the amount of protein and creatinine is determined in a single portion of urine, then the protein/creatinine ratio is used to determine the loss of protein per day.

The method is based on the fact that the rate of excretion of creatinine in the urine is a constant value, does not change during the day. In a healthy person, the normal protein:creatinine ratio in urine is 0.2.

This method eliminates possible errors that may occur when collecting daily urine.

Qualitative samples are more likely than quantitative tests to give false positive or false negative results. Errors occur in connection with taking medications, eating habits, physical activity on the eve of the test.

The interpretation of this qualitative test is given by the visual assessment of turbidity in the test tube in comparison of the test result with the control:

weakly positive reaction is estimated as +;
positive ++;
sharply positive +++.

The Heller ring test is more accurate in assessing the presence of protein in the urine, but does not quantify protein in the urine. Like the sulfosalicylic acid test, the Heller test gives only a rough idea of the amount of protein in urine.

The method allows you to quantify the degree of proteinuria, but it is too laborious, inaccurate, since with a strong dilution, the accuracy of the assessment decreases.

To calculate the protein, you need to multiply the degree of dilution of urine by 0.033 g / l:

1	1	1: 2	0,066
1	2	1: 3	0,099
1	3	1: 4	0,132
1	4	1: 5	0,165
1	5	1: 6	0,198
1	6	1: 7	0,231
1	7	1: 8	0,264
1	8	1: 9	0,297
1	9	1: 10	0,33

The test does not require special conditions, this procedure is easy to do at home. To do this, you need to lower the test strip into the urine for 2 minutes.

The results will be expressed by the number of pluses on the strip, the decoding of which is contained in the table:

Test results corresponding to values up to 30 mg/100 ml are consistent with physiological proteinuria.
Test strip values of 1+ and 2++ indicate significant proteinuria.
Values 3+++, 4++++ are observed in pathological proteinuria caused by kidney disease.

Test strips can only approximately determine the increased protein in the urine. They are not used for accurate diagnosis, and even more so they cannot say what it means.

Do not allow test strips to adequately assess the amount of protein in the urine of pregnant women. A more reliable method of assessment is the determination of protein in daily urine.

Determination of protein in urine using a test strip:

Daily protein in the urine serves as a more accurate diagnosis of the assessment of the functional state of the kidneys. To do this, you need to collect all the urine excreted by the kidneys per day.

Acceptable values for the ratio of protein / creatinine are the data shown in the table:

If you lose more than 3.5 g of protein per day, the condition is called massive proteinuria.

If there is a lot of protein in the urine, a second examination is required after 1 month, then after 3 months, according to the results of which it is established why the norm is exceeded.

The causes of increased protein in the urine are its increased production in the body and disruption of the kidneys, proteinuria is distinguished:

physiological - minor deviations from the norm are caused by physiological processes, are resolved spontaneously;
pathological - changes are caused as a result of a pathological process in the kidneys or other organs of the body, progresses without treatment.

A slight increase in protein can be observed with abundant protein nutrition, mechanical burns, injuries, accompanied by an increase in the production of immunoglobulins.

A mild degree of proteinuria can be caused by physical activity, psycho-emotional stress, and taking certain medications.

Physiological proteinuria refers to an increase in protein in the urine in children in the first days after birth. But already after a week of life, the content of protein in the urine of a child is considered as a deviation from the norm and indicates a developing pathology.

Kidney diseases, infectious diseases are also sometimes accompanied by the appearance of protein in the urine.

Such conditions usually correspond to a mild degree of proteinuria, are transient phenomena, quickly disappear on their own, without requiring special treatment.

More severe conditions, severe proteinuria is noted in the case of:

glomerulonephritis;
diabetes
heart disease;
bladder cancer;
multiple myeloma;
infection, drug-induced injury, polycystic kidney disease;
high blood pressure;
systemic lupus erythematosus;
Goodpasture's syndrome.

Intestinal obstruction, heart failure, hyperthyroidism can cause traces of protein in the urine.

Varieties of proteinuria are classified in several ways. For a qualitative assessment of proteins, you can use the Yaroshevsky classification.

According to the systematics of Yaroshevsky, created in 1971, proteinuria is distinguished:

renal - which includes a violation of glomerular filtration, the release of tubular protein, insufficient readsorption of proteins in the tubules;
prerenal - occurs outside the kidneys, excretion of hemoglobin, proteins that occur in excess in the blood as a result of multiple myeloma;
postrenal - occurs in the area of the urinary tract after the kidneys, the excretion of protein during the destruction of the urinary organs.

To quantify what is happening, the degree of proteinuria is conditionally isolated. It must be remembered that they can easily become more severe without treatment.

The most severe stage of proteinuria develops when more than 3 g of protein is lost per day. Loss of protein from 30 mg to 300 mg per day corresponds to a moderate stage or microalbumnuria. Up to 30 mg of protein in daily urine means a mild degree of proteinuria.

How much protein in urine?

Normally, there is practically no protein in the urine (less than 0.002 g / l). However, under certain conditions, a small amount of protein may appear in the urine of healthy individuals after taking a large amount of protein food, as a result of cooling, with emotional stress, prolonged physical activity (the so-called marching proteinuria).
The appearance of a significant amount of protein in the urine (proteinuria) is a pathology. Proteinuria can be caused by diseases of the kidneys (acute and chronic glomerulonephritis, pyelonephritis, nephropathy of pregnancy, etc.) or urinary tract (inflammation of the bladder, prostate, ureters). Renal proteinuria can be organic (glomerular, tubular and excessive) and functional (feverish proteinuria, orthostatic in adolescents, with overfeeding infants, in newborns). Functional proteinuria is not associated with renal pathology. The daily amount of protein varies in patients from 0.1 to 3.0 g or more. The composition of urine proteins is determined by electrophoresis. The appearance of Bence-Jones protein in the urine is characteristic of multiple myeloma and Waldenström's macroglobulinemia, #223;2 microglobulin in case of damage to the renal tubules.
Normally, there is practically no protein in the urine (less than 0.002 g/l).
The main signs of disease detected in the study of urine.
SG Specific gravity. A decrease in specific gravity indicates a decrease in the ability of the kidneys to concentrate urine and remove toxins from the body, which happens with kidney failure. The increase in specific gravity is associated with a large amount of sugar and salts in the urine. It should be noted that it is impossible to estimate the specific gravity by only one urinalysis, there may be random changes, it is necessary to repeat the urinalysis 1-2 times.

Proteine Protein in the urine - proteinuria. The cause of proteinuria can be damage to the kidneys themselves with nephritis, amyloidosis, damage by poisons. Protein in the urine may also appear due to diseases of the urinary tract (pyelonephritis, cystitis, prostatitis).

Glucose Glucose (sugar) in the urine - glucosuria - is most commonly caused by diabetes. A more rare cause is damage to the renal tubules. It is very alarming if ketone bodies are determined along with sugar in the urine. This happens in severe, misregulated diabetes mellitus and is a harbinger of the most severe complication of diabetes - diabetic coma.

Bilirubin, Urobilinogen Bilirubin and urobilin are determined in the urine in various forms of jaundice.

Erythrocytes Erythrocytes in the urine - hematuria. This happens either when the kidneys themselves are damaged, most often with their inflammation, or in patients with diseases of the urinary tract. If, for example, a stone moves along them, it can injure the mucous membrane, there will be red blood cells in the urine. A decaying kidney tumor can also lead to hematuria.

Leukocytes Leukocytes in the urine - leukocyturia, most often a consequence of inflammatory changes in the urinary tract in patients with pyelonephritis, cystitis. Leukocytes are often determined with inflammation of the female external genital organs, in men - with inflammation of the prostate gland.

Cylindrs Cylinders are peculiar microscopic formations. Hyaline cylinders in the amount of 1-2 can be in a healthy person. They are formed in the renal tubules, these are particles of protein stuck together. But an increase in their number, cylinders of other types (granular, erythrocyte, fatty) always indicate damage to the kidney tissue itself. There are cylinders in inflammatory diseases of the kidneys, metabolic lesions, for example, diabetes mellitus.

Informativity of the method and its limits. The informativeness of a general urine test for recognizing specific kidney diseases is low; additional, more accurate studies are usually required. But this study is very important, especially when conducting preventive studies, as it allows you to identify early signs of kidney disease. It is also known that often kidney diseases are hidden, and only a urine test allows them to be suspected and further necessary examinations to be carried out.
In most laboratories, when testing urine for protein, they first use qualitative reactions that do not detect protein in the urine of a healthy person. If the protein in the urine is detected by qualitative reactions, a quantitative (or semi-quantitative) determination is carried out. At the same time, the features of the methods used, covering a different spectrum of uroproteins, are important. So, when determining protein using 3% sulfosalicylic acid, the amount of protein up to 0.03 g/l is considered normal, while using the pyrogallol method, the limit of normal protein values rises to 0.1 g/l. In this regard, the analysis form must indicate the normal value of the protein for the method used by the laboratory.
When determining the minimum amounts of protein, it is recommended to repeat the analysis; in doubtful cases, the daily loss of protein in the urine should be determined. Normal daily urine contains protein in small quantities. Under physiological conditions, the filtered protein is almost completely reabsorbed by the epithelium of the proximal tubules and its content in the daily amount of urine varies according to different authors from traces to 20-50, 80-100 mg and even up to 150-200 mg. Some authors believe that daily protein excretion in the amount of 30-50 mg/day is the physiological norm for an adult. Others suggest that urinary protein excretion should not exceed 60 mg/m2 of body surface per day, except in the first month of life, when physiological proteinuria may be four times the indicated value.

The general condition for the appearance of proteins in the urine of a healthy person is their sufficiently high concentration in the blood and a molecular weight of not more than 100-200 kDa.
this is not the norm, with your diagnosis this is possible, another thing is that for nephrotic syndrome this is actually a small indicator .. look at the clinic - swelling, pressure, etc. continue taking the prescribed treatment ..
and yet I will say: it should NOT be normal!

Normal indicators: protein is normally found in urine in minimal amounts that are not detected by conventional qualitative reactions. The upper limit of the norm of protein in the urine is 0.033 g / l. If the protein content is higher than this value, then the qualitative tests for protein become positive.

Clinical significance of the definition:

The appearance of protein in the urine is called proteinuria. Proteinuria can be false and renal. Extrarenal proteinuria can be in the presence of impurities of protein origin from the genital organs (vaginitis, urethritis, etc.), while the amount of protein is insignificant - up to 0.01 g / l. Renal proteinuria can be functional (with hypothermia, physical exertion, fever) and organic - with glomerulonephritis, pyelonephritis, nephritis, nephrosis, renal failure. With renal proteinuria, the protein content can be from 0.033 to 10 - 15 g / l, sometimes higher.

quality definition.

Method principle: based on the fact that the protein under the action of inorganic acids coagulates (becomes visible). The degree of turbidity depends on the amount of protein.

Detection of protein in urine with 20% sulfosalicylic acid.

Reagents: 20% solution of sulfosalicylic acid. Equipment: dark background.

Research progress:

2. Pour 2 ml of prepared urine into 2 test tubes of the same diameter. 1 tube - control, 2 - experiment. Add 4 drops of 20% sulfosalicylic acid to the test tube.

3. The result is noted on a dark background.

4. In the presence of protein, the urine in the test tube becomes cloudy.

Qualitative determination of protein in the urine test - strips.

To detect proteinuria, various monotests are used - strips: Albufan, Albustiks, Biofan E and polytests: Triscan, Nonafan, etc.

Quantitation.

Detection of protein in urine by the Roberts-Stolnikov method.

Method principle: based on the fact that the protein under the action of inorganic acids coagulates (becomes visible). The degree of turbidity depends on the amount of protein (i.e. Heller's ring test). At a protein concentration in the urine of 0.033 g/l, a thin, filamentous white ring appears by the end of 3 minutes after layering the urine.

Reagents: 50% nitric acid solution or Roberts reagent (98 parts saturated sodium chloride solution and 2 parts concentrated hydrochloric acid) or Larionova reagent (98 parts saturated sodium chloride solution and 2 parts concentrated nitric acid).

Equipment: dark background.

Research progress:

1. Requirements for urine: urine must be acidic (or slightly acidic) pH, must be transparent, for this urine is centrifuged. Alkaline urine is acidified to a slightly acidic reaction of the medium, using indicator paper for control.

2. Pour 2 ml of 50% nitric acid solution or one of the reagents into the test tube, then carefully layer the same volume of prepared urine along the test tube wall with a pipette

3. The sample is left for 3 minutes

4. After 3 minutes, report the result. The result is noted on a dark background in transmitted light. If the ring is wide, compact, then the urine is diluted with distilled water and again layered on the reagent.

5. Urine is diluted until a thin thread-like ring forms after 3 minutes.

C \u003d 0.033 g / l x the degree of dilution.

For the clinic, both qualitative and quantitative determination of protein in the urine is important.

Qualitative tests for the determination of protein in the urine
More than 100 reactions for the qualitative determination of protein in urine have been proposed. Most of them are based on protein precipitation by physical (heating) or chemical means. The presence of protein is proved by the appearance of turbidity.

Colorimetric dry samples are also of interest.

Only the most important samples for practice will be described below.

Sample with sulfosalicylic acid. To several milliliters of urine, add 2-4 drops of a 20% solution of sulfosalicylic acid. With a positive reaction, turbidity appears. The result is denoted by the terms: opalescence, weakly positive, positive or strongly positive reaction. The sulfosalicylic acid test is one of the most sensitive tests for determining protein in the urine. It detects even the most insignificant pathological increases in protein in the urine. Thanks to a simple technique, this test has found wide application.

Aseptol test. Aseptol is a substitute for sulfosalicylic acid. It can be prepared from materials available in any laboratory (phenol and sulfuric acid). A 20% solution of aseptol is used as a reagent. The test is carried out as follows: in a test tube containing 2-3 ml of urine, add 0.5-1 ml of aseptol solution to the bottom. If a white ring of coagulated protein forms at the interface between the two fluids, the test is positive.

Geller test. Under a few milliliters of urine, add 1-2 ml of 30% nitric acid (sp. weight 1.20). If a white ring appears at the interface of both liquids, the sample is positive. The reaction becomes positive if the protein is more than 3.3 mg%. Sometimes a white ring is obtained in the presence of large amounts of urates. Unlike the protein ring, the urate ring does not appear at the border between both fluids, but a little higher. Larionova suggests that instead of 30% nitric acid, use as a reagent a 1% solution of nitric acid in a saturated solution of common salt; this gives a great saving of nitric acid.

Test with ferruginous potassium and acetic acid. This reaction makes it possible to distinguish serum proteins from nucleoalbumins.

Equal amounts of urine are poured into two test tubes. A few drops of a 30% acetic acid solution are added to one of them. If there is haze compared to the control tube, the urine contains nucleoalbumin. If turbidity does not appear, the contents of both tubes are mixed and again divided into two parts. To one of the two test tubes, add a few drops (an excess can turn a positive sample into a negative one) of a 10% solution of yellow blood salt (potassium ferricyanide). In the presence of whey proteins, turbidity is obtained.

With concentrated urine containing large amounts of uric acid and urates, a test with potassium ferricyanide and acetic acid should be performed after preliminary dilution (2-3 times) of urine with water. Otherwise, cloudiness caused by settled uric acid may occur.

This is especially important when examining the urine of infants containing a lot of uric acid and urates.

Of the other qualitative tests for protein in the urine, based on the precipitation of proteins, the following have been used: the boiling test, the tests of Esbach, Purdy, Roberts, Almen, Balloni, Buro, Claudius, Corso, Dome, Goodmann-Suzanne, Jollet, Exton, Kamlet, Kobuladze , Liliendal-Petersen, Polacci, Pons, Spiegler, Tanre, Thiele, Brown, Tsushiya, etc.

In the production of quality urine protein samples based on protein precipitation, the following general rules must be observed, the violation of which leads to significant errors in the analysis.

1. The urine to be tested must be acidic. In an alkaline reaction, the urine is slightly acidified with acetic acid. Producing an alkaline urine sample when acid is used as a reagent may result in neutralization of the acid and a negative result if positive. This is especially true for the sulfosalicylic acid sample, since the acid is added in very small amounts and can be easily neutralized.

2. The urine to be tested must be clear.

3. Samples for the determination of protein in the urine should always be made in two test tubes, one of which serves as a control. Without a control tube, you may not notice slight turbidity in the reactions.

4. The amount of added acid in the samples should not be too large. A large amount of acid can lead to the formation of soluble acidalbumins and to the transformation of a positive sample into a negative one.

Deserve great attention, due to their simple technique, colorimetric dry samples. These tests use the influence that a protein has on the color of an indicator in a buffer solution (the so-called indicator protein error). A filter paper strip impregnated with acid citrate buffer and bromophenol blue as an indicator is immersed for a short time in the urine. The test is positive if a blue-green color is obtained. By comparing the color intensity with color paper standards, tentative and quantitative conclusions can also be drawn. Indicator papers are sold in reams of appropriate color standards, similar to universal indicator papers.

Methods for quantitative determination of protein in urine
Many methods have been proposed for the quantitative determination of protein in urine. Precise quantitative methods for the determination of proteins in biological material have not found wide application in the determination of protein in urine, due to complex and time-consuming techniques. Volumetric methods, especially the Esbach method, are widely used. They are very simple, but, unfortunately, they are not very accurate. The methods of the Brandberg-Stolnikov group are also convenient for the clinic, giving more accurate results than volumetric methods, with a relatively simple technique. In the presence of a photometer or nephelometer, nephelometric methods are also convenient.

Esbach method. It was proposed by the Parisian doctor Esbach in 1874. Urine and a reagent are poured into a special test tube (Esbach's albuminometer). The test tube is corked with a rubber stopper, thoroughly stirred (without beating!) and left in an upright position until the next day. They report the division, to which the column of protein sediment reaches. The number found shows the protein content. It is very important with the Esbach method that the urine is acidic. Alkaline urine can neutralize the acidic constituents of the reagent and prevent the precipitation of proteins.

Advantages of the method: it is simple and convenient in practice.

Disadvantages: the method is inaccurate, the result is obtained after 24 - 48 hours.

Brandberg-Stolnikov method. It is based on a qualitative Geller test. The Geller test can be used for quantitation as it gives a positive result above 3.3 mg% protein. This is the limiting protein concentration below which the sample becomes negative.

Erlich and Althausen modification. Soviet scientists S. L. Erlikh and A. Ya. Althausen modified the Brandberg-Stolnikov method, indicating the possibility of simplifying the study and saving time in its production.

The first simplification is related to the time of the appearance of the ring. The exact time of its appearance is determined, without necessarily adhering to the 2nd and 3rd minutes.

The second simplification makes it possible to establish what dilution should be made. The authors proved that the required dilution can be approximately determined by the type of the resulting ring. They distinguish filiform, wide
and a compact ring.

Of the nephelometric methods, it deserves to be noted Kingsberry and Clark method. 2.5 ml of filtered urine is poured into a small graduated cylinder, replenished with a 3% aqueous solution of sulfosalicylic acid to 10 ml. Stir thoroughly and after 5 minutes photometer in a 1 cm cuvette, with a yellow filter, using water as a compensation liquid. With a Pulfrich photometer, the extinction found, multiplied by 2.5, gives the amount of protein in %o. In the case when the extinction index is higher than 1.0, the urine is pre-diluted 2 times, 4 times or even more.

In order to have a clear idea of the amount of proteins excreted in the urine, it is necessary to determine not only their concentration in a separate portion of urine, but also their total daily amount. To do this, collect the patient's urine for 24 hours, measure its volume in milliliters and determine the protein concentration in a portion of daily urine in g%. The amount of proteins excreted in the urine in 24 hours is determined depending on the daily amount of urine in grams.

Clinical significance of protein in urine

Human urine normally contains minimal amounts of protein that cannot be ascertained by routine qualitative urine protein tests. The excretion of large amounts of protein, in which ordinary qualitative tests for protein in the urine become positive, is an abnormal phenomenon called proteinuria. Proteinuria is physiological only in a newborn, in the first 4-10 days after birth. The commonly used name albuminuria is incorrect, because not only albumins, but also other types of proteins (globulins, etc.) are excreted in the urine.

Proteinuria, as a diagnostic symptom, was discovered in 1770 by Cotugno.

The most important functional renal proteinuria in children are as follows:

1. Physiological proteinuria of the newborn. It occurs in most newborns and has no adverse significance. It is explained by a weak kidney filter, damage at birth, or loss of fluids in the first days of life. Physiological proteinuria disappears on the 4-10th day after birth (later in premature babies). The amount of protein is small. It is nucleoalbumin.

Long-term neonatal albuminuria may be a symptom of congenital lues.

2. Stroke albuminuria. They are caused by exceeding the threshold of normal irritability of the renal filter by significant mechanical, thermal, chemical, mental and other irritations - loss of fluid in infants (dehydrational proteinuria), cold bathing, abundant, protein-rich food (alimentary proteinuria), palpation of the kidney (palpatory albuminuria), physical overwork, fear, etc.

Stroke albuminuria appears more easily in children at an early age than in older children and adults, since the kidneys of an infant and a small child are more easily irritated. Dehydration albuminuria (malnutrition, hydrolability, toxicosis, diarrhea, vomiting) is especially often observed in infants.

Stroke albuminuria is benign. They disappear immediately after the elimination of the causes that cause them. In a deposit sometimes there are single leukocytes, cylinders and erythrocytes. The protein is most often nucleoalbumin.

3. Orthostatic proteinuria. This condition is typical for children of preschool and school age. It occurs on the basis of vasomotor disorders of the blood supply to the kidney. Typical for orthostatic albuminuria (hence its name) is that it appears only when the child is standing, when the spine is in a lordotic position. In the supine position, it disappears. Nucleoalbumin is released. In doubtful cases, you can resort to orthostatic experience, which is as follows: in the evening, an hour before going to bed, the child empties the bladder; in the morning, getting out of bed, he urinates again. This urine does not contain protein. Then the child is put on his knees for 15-30 minutes with a stick behind his back, between the bent elbows of both hands. A position of lordosis is created, which leads to the release of protein, without changes in the sediment.

With orthostatic albuminuria, 8-10 g of protein can be secreted per day.

The most important clinical significance among all proteinuria are organic renal proteinuria. They are caused by organic diseases of the kidneys (nephritis, nephrosis, nephrosclerosis). Proteinuria is one of the most important and best known symptoms of organic kidney disease.

1. In acute and chronic glomerulonephritis, proteinuria occurs regularly. The amount of protein is moderate, and there is no parallel between the degree of proteinuria and the severity of the disease. In contrast, chronic and more severe nephritis often occurs with less protein than acute ones. After acute nephritis, sometimes for a long time (years), small amounts of protein in the urine are established, which do not have pathological significance ("residual albuminuria"). We should not forget that "nephritis without proteinuria" can also occur. Sometimes the protein is found in one portion of the urine, but not in the other. The ratio of albumins to globulins in acute nephritis is low, and in chronic nephritis it is higher.

2. With nephrosclerosis, the amount of protein in the urine is very small, often there are forms of the disease without protein in the urine.

3. Of all renal diseases, nephrosis occurs with the most pronounced proteinuria.

4. In infectious and toxic conditions, so-called febrile and toxic proteinuria occur. These are acute nephroses, in which the amount of protein is small. This group also includes proteinuria in convulsive conditions (convulsions), hyperthyroidism, jaundice, intussusception, enterocolitis, burns, severe anemia, etc. These albuminuria are benign and pass quickly (transient albuminuria).

5. With stagnation of blood in the kidneys, the so-called congestive albuminuria occurs, which is characteristic of heart patients in the stage of decompensation. It is also found in ascites and tumors of the abdomen.

With febrile, toxic and congestive albuminuria, the increased permeability of the renal filter is especially pronounced. According to some authors, many of these proteinuria occur without organic damage to the kidney parenchyma.

Extrarenal albuminuria are usually caused by protein impurities (secretions, decayed cells), which are excreted by diseased urinary tract and genital organs. Extrarenal albuminuria is more common due to cystopyelitis (pyuria), less often due to vulvovaginitis, calculi and tumors of the urinary tract.

With extrarenal albuminuria, a large number of leukocytes and bacteria are found in the sediment. Renal elements are almost never found. The amount of protein is small. Filtered or centrifuged urine usually does not test positive for protein.

In those recovering from pyelitis, albuminuria disappears after bacteriuria and pyuria.

It should be emphasized as a characteristic phenomenon that organic renal diseases appear extremely rarely in early childhood, and therefore organic proteinuria is also rare. Of these, there are mainly febrile and toxic. In contrast to organic proteinuria, stroke albuminuria is very common in children at an early age.

In older children, organic proteinuria is more often functional. In general, with age, functional proteinuria is less common, and organic more often.

Electrophoretic studies of proteins in urine

A number of authors use the electrophoretic method to study proteins in the urine (uroproteins). It can be seen from the obtained electrophoregrams that they have the same qualitative composition as plasma proteins. This indicates that the proteins in the urine are derived from plasma proteins.

R-you 50% solution of nitrogen to-you or p-larionic. The course of determination: a number of test tubes are placed in a tripod and 1 ml of nitric acid solution is poured, add 1 ml of urine, layered on the reagent and time is recorded, when a ring appears, we record the time of the appearance of the ring. If the ring is wide, dilute urine.

4. Determination of protein concentration in urine with 3% sulfosalicylic acid.

Solutions: 3% sc, sodium chloride 9%, pp albumin 10%. Progress of determination: 1.25 ml of clear urine are placed in two measured centrifuge tubes "O" - experience and "K" - control. 3.75 ml of a 3% solution of sulfosalicylic acid is added to the experimental one, and 3.75 ml of a 0.9% solution of sodium chloride is added to the control. Leave for 5 minutes, and then photometry on FEC at a wavelength of 590 - 650 nm (orange or red light filter) in a cuvette with a layer thickness of 5 mm experiment against control. The calculation is carried out according to the calibration graph or table. Method principle is based on the fact that protein with sulfosalicylic acid gives turbidity, the intensity of which is directly proportional to the concentration of the protein.

5. detection of glucose in the urine, the Gaines-Akimov test. Principle: Glucose, when heated in an alkaline medium, reduces copper dihydroxide (yellow) to copper monohydroxide (orange-red). Reagent preparation: 1) 13.3 g chem. pure crystalline copper sulfate (CuSO 4 . 5 H 2 O) solution. in 400 ml of water. 2) 50 g of caustic soda is dissolved in 400 ml of water. 3) 15 g of pure glycerin is diluted in 200 ml of water. Mix the first and second solutions and immediately add the third. Reagent racks. Definition progress: 1 drop of urine and 9 drops of reagent are added to the test tube and boiled in a water bath for 1-2 minutes. Positive test: yellow or orange color of liquid or precipitate.

6. Qualitative determination of glucose in urine by the glucose oxidase method. Method principle: glucose is oxidized in the presence of glucose oxidase, according to the reaction: Glucose + O 2 with glyconolant + H 2 O 2. The formed peroxide H under the action of peroxidase oxidizes the substrate with the formation of a colored product.

Pour in and incubate for 15 minutes at 37 0 C. Look at the CPK, 5mm cuvette.

Then, calculations are made according to the formula: С op = Ext op . Cst/ Ect st.

7. Detection of ketone bodies in urine by Lestrade test. On the glass slide is applied (at the tip of the scalpel) a powder or a tablet of Lestrade solution, and 2-3 drops of urine are applied to it. In the presence of ketone bodies, a pink to purple color will appear. The sample is evaluated on a white background.

8. Detection of blood pigment in the urine by a test with a 5% alcohol solution of amidopyrine.

1.5% alcohol solution of amidopyrine (0.5 g of amidopyrine is dissolved in 10 ml of alcohol 96%) 2.3% solution of hydrogen peroxide 1.5 g of hydropyrite is dissolved in 50 ml of water) -ether extract or shaken unfiltered urine. add 8-10 drops of 5% amidopyrine solution and 8-10 drops of 3% hydrogen peroxide solution; take into account the result no later than 2-3 minutes. The sample is considered positive in the presence of gray-violet staining.

Detection of urobilin in the urine with a Neubauer test.

It is based on the color reaction of urobilinogen with Erlich's reagent, which consists of 2 g of paradimethylaminobenaldehyde and 100 ml of hydrochloric acid solution (200 g.l). 1 ml of urine and 1 ml of solution. The appearance of red color in the first 30 seconds indicates an increase in urobilinogen content. Normally, color appears later or is completely absent. When urine stands, urobilinogen turns into urobilin and the test may be false negative. cannot be heated, because by-product complex compounds, aldehyde with porphyrins, indole and drugs can be formed.

Detection of bilirubin in the urine with a Rosin test.

Alcoholic solution of iodine (10g.l): 1 g of crystalline iodine is dissolved in a cylinder with a capacity of 100 ml in 20-30 ml of 96 g of rectified alcohol, and then topped up with alcohol to the mark. The course of determination. Pour into a chemical test tube 4-5 ml of the test urine and carefully layer an alcohol solution of iodine on it (if the urine has a low relative density, then it should be layered on an alcohol solution of iodine). antipyrine, as well as when there is a blood pigment in the urine, the test turns out to be positive). In a healthy person, this test is negative.

Urine examination by dry chemistry (monopolytests).

Principle. The method is based on the effect exerted by the protein on the color of the indicator in the buffer solution, as a result of which the dye changes color from yellow to blue.

When testing for the presence of protein in the urine and determining the pH using indicator paper, it is recommended that the following guidelines be followed:

Collect urine in a thoroughly washed dish.
Use freshly collected, preservative-free urine.
Close the case carefully after removing the required number of indicator strips of paper from it.
Do not grab the indicator zones with your fingers.
Use only within the expiration date stated on the label.
Observe the rules for storing indicator paper.
Evaluate the results in accordance with the instructions available in the instructions.

Performing a urine test on a dry urine chemistry analyzer.

Definition progress. A strip of indicator paper is removed from the case and immersed in the test urine so that both indicator zones are moistened simultaneously. After 2-3 seconds, the strip is placed on a white glass plate. Immediately carry out a pH assessment using the color scale printed on the pencil case. The pH value on the color scale corresponds to 6.0 (or less); 7.0; 8.0; 9.0.

Urine preparation, preparation of preparations from urine sediment by microscopic examination in an approximate way.

Microscopic examination of the urine sediment is carried out by an approximate method in the general analysis and quantitative calculation of shaped elements for a more accurate assessment of the degree of leukocyturia and hematuria.

Rules for the preparation of urine sediment for microscopy.

The first morning portion of urine is subject to microscopic examination.

After preliminary mixing, 10 ml of urine is taken, centrifuged for 10 minutes at 1500 rpm.

Then the centrifuge tube with urine is overturned with a sharp movement, the supernatant is quickly poured into an empty jar.

Stir, place a drop on a glass slide and carefully cover with a coverslip.

If the precipitate consists of several layers, then prepare the preparation, and then centrifuge again and prepare preparations from each layer separately.

In the absence of a sediment visible to the eye, a drop of urine is applied to a glass slide and microscopically.

At the beginning, the material is examined at low magnification (eyepiece 7-10, objective 8), while the condenser is lowered, the diaphragm is slightly narrowed, then the preparation is studied in detail at high magnification (eyepiece 10.7; objective 40).

14.Quantitative study of urine sediment according to Nechiporenko.

The method is used for latent sluggish inflammatory processes (pyelonephritis, glomerulonephritis), latent pyuria. To study the pathological process in dynamics. To evaluate the effectiveness of the treatment. Advantages of the method: technically simple, does not require a large amount of urine and is long lasting. its storage, is used in outpatient practice. Mandatory conditions: morning urine, medium portion, acidic solution (in alkaline there may be a partial breakdown of cellular elements). 1. Urine is mixed. 2. 10 ml of urine is placed in a measuring centrifuge tube and centrifuged for 10 minutes at 1500 rpm. 3. After centrif. sucking Pipette the top of the liquid, leave. exactly 1 ml of sediment. 4. The precipitate is thoroughly mixed and the Goryaev chamber is filled. 5. After 3-5 minutes after filling, they begin to count the shaped elements. 6. Counting of leukocytes, Er, cylinders with an eyepiece 15 objective 8 when lowered. condenser, in 100 large chamber squares. White blood cells, Er are counted separately, cylinders (counting at least 4 Goryaev chambers) are excreted. arith. X \u003d A x 0.25x 10 6 / l. Norm: lake. 2-4x 10 6 /l, Er up to 1 x 10 6 /l, cylinders up to 0.02 x 10 6 /l (one for 4 chambers). In children: leuk. up to 2-4x 10 6 /l, Er up to 0.75 x 10 6 /l, cylinders up to 0.02 x 10 6 /l.

15. Urinalysis according to Zimnitsky

This test determines the ability of the kidneys to concentrate. and dilute urine. Essence of the sample in the dynamic determination of the relative density and quantity of urine in three hours in portions during the day. Conducting a test: after emptying the bladder at 6 o'clock in the morning into the toilet, the patient collects urine every three hours in separate jars during the day. Only 8 servings. Research progress: 1. Delivery. Urine is arranged by the hour and in each portion determine the amount and relative density. 2. Compare the daily amount of urine and the amount of liquid drunk to determine. % of its excretion. 3. Calculate day and night diuresis, summarize, get daily diuresis. 4. Set the fluctuation range of quantity and relative. urine density per day i.e. what is the difference between the smallest portion and the largest. Show. health tests. people: 1. Daily diuresis 800-1500 ml. 2. Day diuresis significantly prevails over night. 3. Fluctuations in the volume of urine in individual portions are significant (from 50 to 400 ml). 4. Fluctuations p from 1.003 to 1.028, should be more than 0.008. With func. kidney failure: hypostenuria, hypoisostenuria, isostenuria, hyperstenuria, oliguria, anuria, nocturia.

16. Description of the general properties of feces.

Normally, feces consist of products of secretion and excretion of the digestive tract, residues of undigested or partially digested foods, and microbial flora. The amount of feces is 100-150 g. The consistency is dense. The shape is cylindrical. The smell is fecal normal. Brown color. R-tion is neutral, slightly alkaline or slightly acidic (pH 6.5-7.0-7.5). Mucus is absent. Blood is absent. The remains of undigested food are absent.

Instruction

Qualitative methods for the determination of protein in urine: Heller method, 20% sulfosalicylic acid test, boiling test, etc. Semi-quantitative methods: use of diagnostic test strips to determine protein in urine, Brandberg-Roberts-Stolnikov method. Quantitative methods: turbidimetric and colorimetric.

Determination of protein in the daily urine at a concentration of 0.033 g/liter or more is a pathology. As a rule, in the morning portion of urine, the protein concentration does not exceed 0.002 g / l, and in the daily urine protein concentration is not more than 50-150 mg of protein.

Sources:

determination of protein in urine

Urine is a product of human metabolism. It is formed during the filtration of blood in the kidneys, which is why the composition of urine gives a clear description of the state of the human body.

Urine is a complex solution of more than 150 compounds. Some specific substances, for example, acetone, bile acids, protein, glucose, may be present only in certain diseases.

To control human health, first of all, it is necessary to determine the amount of urine. The norm is the formation of 1-1.8 liters of urine per day. When more than 2 liters of urine is excreted, this is a sign of a possible disruption in the functioning of the kidneys, diabetes mellitus and a number of other diseases. If less than 0.5 liters of urine is formed per day, there is a blockage of the ureter or bladder.

urine color

The color of urine excreted depends on many factors, so it can vary, ranging from light yellow to orange. The presence of certain shades can be affected by certain foods, as well as medications taken by a person.

After taking medications, urine may become stained and acquire a reddish tint. If a person is actively moving, while he releases a large amount of sweat, the urine will have an intense yellow color, as when taking drugs such as Nitroxoline or Biomycin.

If a person has not taken any coloring foods and medicines, but the color of his urine is different from usual, one can suspect the presence of a disease in the body. For example, in diseases of the liver, urine will have a dark yellow or greenish color.

The presence of blood in the excreted urine clearly indicates the presence of a stone in or renal bleeding, if pain is also observed.

If urination is difficult, this may indicate an inflammatory process caused by an infection in the bladder. But dirty and cloudy urine indicates severe kidney disease.

Protein in the urine

There is no protein in the blood of a person or its amount is so small that it cannot be determined using laboratory tests. If a protein is detected in the urine, it is necessary to conduct repeated tests, since it can be present when a person wakes up in the morning, as well as after hard physical work or exercise in athletes.

To determine visually whether protein is present in the urine or not is 100% impossible. One can only guess when there is a large amount of whitish flakes in the urine.

If the protein in the urine is repeatedly detected, this indicates the presence of some kind of kidney disease. The inflammatory processes occurring in them provoke a slight increase in the amount of protein. If more than 2 grams are excreted in the urine, this is an alarm signal.

Pyelonephritis is an inflammatory disease that affects the renal pelvis, calyces, and parenchyma. In most cases, inflammation is caused by a bacterial infection. Full recovery is possible only with timely diagnosis, therefore, when symptoms appear, a thorough examination is necessary.