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URINALYSIS.

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Presentation on theme: "URINALYSIS."— Presentation transcript:

1 URINALYSIS

2 URINALYSIS :- THE CHEMICAL EXAMINATION OF URINE

3 Introduction

4 CHEMICAL EXAMINATION OF URINE
the routine urinalysis includes chemical testing for: pH ,protein, glucose, ketones, occult blood, bilirubin, urobilinogen, nitrite, leukocyte esterase, and strip test method for specific gravity

5 Compound of urine :- nitrogen compound of urine :- 1-urea 2- uric acid 3- creatinine . Non nitrogen compound of urine :- 1-vitamins 2-amino acids 3-chloride 4-potassium 5-phosphate 6-sulfate 7- calcium 8-magnesium

6 THE CHEMICAL EXAMINATION OF URINE
REAGENT STRIPS :- Reagent strip consist of chemical-impregnated absorbent pads attached to the plastic strip. A color- producing chemical reaction takes place when the absorbent pad comes in contact with urine.

7 Reagent strip is the most common in chemical examination of
urine………why??????? simple, rapid means for performing 10 medically significant chemical analyses, including pH, protein, glucose, ketones, blood, bilirubin, urobilinogen, nitrite, specific gravity UrinCheck™ HealthScreen-10 Reagent Strips Procedu…: http://youtu.be/CPu-N9YnJGo UrinCheck™ HealthScreen-10 Reagent Strips Procedu…: http://youtu.be/CPu-N9YnJGo video for reagent strips proceduer :

8 Sources of Error for reagent srtip:
1) Testing cold specimens - would result in a slowing down of reactions; test specimens when fresh or bring them to RT before testing 2) Inadequate mixing of specimen - could result in false reduced or negative reactions to blood and leukocyte tests; mix specimens well before dipping 3) Over-dipping of reagent strip - will result in leaching of reagents out of pads; briefly, but completely dip the reagent strip into the urine

9 THE CHEMICAL EXAMINATION OF URINE

10 pH the kidneys are the major regulators of the acid-base content in the body. - A healthy individual will usually produce first morning specimen with a slightly acidic pH of 5.0 to 6.0, the pH of normal random samples can range from 5.0 to 8.0. principle of test is based on the double-indicator system of methyl red and bromothymol blue.

11 CLINICAL SIGNIFICANCE OF URINE pH
A highly acidic urine pH occurs in: Acidosis Uncontrolled diabetes Diarrhea Starvation and dehydration Respiratory diseases in which carbon dioxide retention occurs and acidosis develops A highly alkaline urine occurs in: Urinary tract obstruction Pyloric obstruction Salicylate intoxication Renal tubular acidosis Chronic renal failure Respiratory diseases that involve hyperventilation (blowing off carbon dioxide and the development of alkalosis)

12 pH Source of error: A number of preanalytical variables can affect urine pH. Bacterial overgrowth in a specimen standing at room temperature will often lead to a higher pH due to the conversion of urea to ammonia. Diets with a high content of vegetables and citrus fruits may produce an alkaline pH. source of error

13 Protein As a routine chemical tests performed on urine the most indicative of renal disease is the protein determination. The presence of proteinuria is often associated with early renal disease. Normal urine contains very little protein; usually, less than 10 mg/dl or 150 mg per 24 hours is excreted.

14 Protein Benign proteinuria is usually transient and can be produced by conditions such as: exposure to cold, strenuous exercise, high fever, dehydration, and in the acute phase of sever illnesses. Negative Trace + (30 mg/dL) ++ (100 mg/dL) +++ (300 mg/dL) ++++ (2000 mg/dL)

15 Protein Test :- HEAT & ACETIC ACID TEST. Principle:-
proteins are denatured & coagulated on heating to give white cloud precipitate. Method:- take 2/3 of test tube with urine, heat only the upper part keeping lower part as control. Presence of phosphates, carbonates, proteins gives a white cloud formation. Add acetic acid 1-2 drops, if the cloud persists it indicates it is protein(acetic acid dissolves the carbonates/phosphates)

16 Sourse :http://medicine-science-and-more

17 Bence Jones proteins Bence Jones protein is a monoclonal globulin protein. Finding this protein in blood and urine is often suggestive of multiple myeloma or Waldenstrom's macroglobulinemia. Test:- Thermal method(waterbath): Proteins has unusual property of precipitating at ( c) & then dissolving when the urine is brought to boiling(1000c) & reappears when the urine is cooled. clinical sign:The Bence-Jones protein urine test is used mainly to diagnose and monitor multiple myeloma, a blood cancer Source of error of protein:- 1- increase of Exercise . 2- increase eating of red meat….why??

18 Glucose Glucose test is the most frequent chemical analysis performed on urine because of its value in the detection and monitoring of diabetes mellitus. principle of test is based on a double sequential enzyme reaction :- -Two very different tests are utilized by laboratories to measure urinary glucose, the glucose oxidase procedure provides a specific test for glucose. Negative Trace (100 mg/dL) + (250 mg/dL) ++ (500 mg/dL) +++ (1000 mg/dL) ++++ (2000+ mg/dL) CLINICAL SIGNIFICANCE OF URINE GLUCOSE: Diabetes mellitus. Renal glycosuria. SOURCE OF Error : 1- take a lot of diet have sugar 2-Drug and some disease that cause increase sugar

19 sugar Test:- BENEDICT’S TEST(semi quantitative) Principle:-
benedict’s reagent contains cuso4.In the presence of reducing sugars cupric ions are converted to cuprous oxide which is hastened by heating, to give the color. Method:- take 5ml of benedict’s reagent in a test tube, add 8drops of urine. Boil the mixture.

20 KETONES The term ketones represents; three intermediate products of fat metabolism, namely, acetone, acetoacetic acid, and beta-hydroxybutyric acid. The three ketone bodies are not present in equal amounts in urine. principle of test is based on the reaction of acetoacetic acid with sodium nitroprusside in a strongly basic medium . Acetoacetic Acid + Nitroprusside > Colored Complex CLINICAL SIGNIFICANCE OF URINE KETONES: Diabetic ketoacidosis Prolonged fasting

21 KETONES TEST:- Rothera’s test. Principle:-
acetone & acetoacetic acid react with sodium nitroprusside in the presence of alkali to produce purple colour. Method:- - take 5ml of urine in a test tube & saturate it with ammonium sulphate. Then add one crystal of sodium nitroprusside. Then gently add 0.5ml of liquor ammonia along the sides of the test tube. - Change in colour indicates + test

22 KETONES Negative Trace (5 mg/dL) + (15 mg/dL) ++ (40 mg/dL)

23 BLOOD OCCULT BLOOD :- The term “occult” means “hidden,” and the methods used to test for blood in the urine are capable of detecting even minute amounts not visualized macroscopically. Another reason for this title is that these procedures actually detect the free hemoglobin from lysed red blood cells (RBCs). principle of test is based on the pseudoperoxidase action of hemoglobin and erythrocytes which catalyzes the reaction of 3, 3’, 5, 5’-tetramethyl-benzidine and buffered organic peroxide . CLINICAL SIGNIFICANCE OF URINE BLOOD :- Hematuria Hemoglobinuria Myoglobinuria

24 BLOOD TEST:- BENZIDINE TEST Principle:-
Principle-The peroxidase activity of hemoglobin decomposes hydrogen peroxide releasing nascent oxygen which in turn oxidizes benzidine to give blue color. Method:- - mix 2ml of benzidine solution with 2ml of hydrogen peroxide in a test tube. Take 2ml of urine & add 2ml of above mixture. A blue color indicates + reaction.

25 BLOOD

26 BILIRUBIN AND UROBILINOGEN
BILIRUBIN AND UROBILINOGEN:- Bilirubin is formed from the breakdown of hemoglobin in the reticuloendothelial system. It is then bound to albumin and transported through the blood to the liver. In the intestines, bacterial enzymes convert bilirubin, through a group of intermediate compounds, to several related compounds which are collectively referred to as Urobilinogen .

27 BILIRUBIN SCREENING TESTS FOR BILIRUBIN (BILE) Bilirubin can be detected in the urine before other clinical symptoms are present or recognizable Negative + (weak) ++ (moderate) +++ (strong) CLINICAL SIGNIFICANCE OF URINE bilirubin :- Increased direct bilirubin (correlates with urobilinogen and serum bilirubin). TEST:- Ictotest (more sensitive tablet version of same assay) Serum test for total and direct bilirubin is more informative

28 UROBILINOGEN SCREENING TESTS FOR UROBILINOGEN:-
0.2 mg/dL 1 mg/dL 2 mg/dL 4 mg/dL 8 mg/dL SCREENING TESTS FOR UROBILINOGEN:- Screening for urobilinogen is useful in the diagnosis of liver function disorders. FOAM TEST:- the urine is a yellowish-brown or greenish-yellow color and bilirubin is suspected, shake the urine. If a yellow or greenish-yellow foam develops, then bilirubin is most likely present. Bilirubin alters the surface tension of urine and foam will develop after shaking. REAGENT TEST STRIPS:- Screening tests for urobilinogen are based on the Ehrlich Aldehyde Reaction: p-Dimethylaminobenzaldehyde + urobiligen = red-colored azo dye

29 UROBILINOGEN CLINICAL SIGNIFICANCE OF URINE BLOOD :-
High: increased hepatic processing of bilirubin . Low: bile obstruction .

30 LEUKOCYTE ESTERASE White blood cells can be present in any body fluid depending on a cause for their presence. The most common white blood cell seen in a urine sample is the neutrophil, which is normally present in low numbers. Increased numbers of neutrophils usually indicate the presence of a urinary tract infection; and their presence is indicated by a positive leukocyte esterase test. Screening for urinary tract infections also includes evaluation of pH, protein, and nitrite A positive reaction produces a lavender to purple color with a reporting range of values from trace to large. Values reflecting cell numbers from negative to 500 may be reported. These results may not correlate with the numbers of neutrophils seen during microscopic examination. False-Positive Results: Strong oxidizing agents cause a false-positive leukocyte esterase result

31 NITRITE The nitrite test:
is a rapid, indirect method for the early detection of significant and symptomatic bacteriuria.Common organisms that can cause urinary tract infections, such as Escherichia coli,Enterobacter, Citrobacter, Klebsiella,and Proteus species, produce enzymes that reduce urinary nitrate to nitrite. For this to occur, the urine must haveincubated in the bladder for a minimum of 4 hours. Hence,the first morning urine is the specimen of choice. Nitrite results are read at 30 or 60 seconds, depending on the manufacturer Pink spots or pink edges should not be considered a positive result. If the uniform pink color is very light, it may best be seen by placing the strip against a white background. The test is reported as positive or negative.

32 Source of error of nitrite in urine:
False-Positive Results The urine should be tested shortly after being voided, because if the urine is allowed to stand at room temperature for several hours, organisms may grow in the specimen and generate nitrite.9,10,48 Results may be misinterpreted as positive in urines that appear red or contain phenazopyridine and other substances that turn red in acid.5,11 False-Negative Results The sensitivity of the test is reduced in urine with a high specific gravity or elevated level of ascorbic acid.9,11 A negative test should never be interpreted as indicating the absence of bacterial infection.

33 NITRITE Interpretation:
Nitrite results are read at 30 or 60 seconds, depending on the manufacturer. Any degree of uniform pink color should be interpreted as a positive nitrite test suggesting the presence of 105 or more organisms per milliliter. The color development is not proportional to the number of bacteria present. Pink spots or pink edges should not be considered a positive result.2,5,9,11 If the uniform pink color is very light, it may best be seen by placing the strip against a white background. The test is reported as positive or negative

34 CLINICAL SIGNIFICANCE for nitrite:
Early detection of bacteria is important in order to prevent cystitis from developing into inflammation or infection involving the kidney and renal pelvis. The nitrite portion of the test strip can be used to screen individuals who are at risk for developing urinary tract infections, such as diabetics, persons with recurrent infections, or pregnant women. The test is also useful in evaluating the success of antibiotic therapy that is used to treat a bladder infection.

35 Thanks for watching our presentation

36 الدكتور/محمد الختاتنة
إعداد الطلاب: -مشعل محمد الشريف -مشار ي سلمان الخيبري -سالم داحش العمري -سعد داحش العمري Multistix© 10 SG reagent strips [Color chart]. Tarrytown, NY: Bayer Corporation Diagnostics Division, 1996. 2. Multistix© 10 SG reagent strips [Package insert]. Tarrytown, NY: Siemens Healthcare Diagnostics Inc, 2008. 3. de Wardener HE. The Kidney. 3rd Ed. Boston: Little, Brown & Co, 1967. 4. James GP, Bee DE, Fuller JB. Accuracy and precision of urinary pH determinations using two commercially available dipsticks. Am J. Clin Pathol 1978;70:368–374. 5. Chemistrip© reagent strips [Package insert]. Indianapolis, IN: Roche Diagnostics Corporation, 1999. 6. James JA. Proteinuria and hematuria in children: diagnosis and assessment. Pediatr Clin North Am 1976;23:807–816. 7. Baker FJ, Silverton RE, Luckcock ED. Introduction to Medical Laboratory Technology. 4th Ed. Washington: Butterworth & Co, 1966. 8. Free HM. Modern Urine Chemistry Manual. Tarrytown, NY: Bayer 9. AimStick [package insert]. San Antonio, TX: Germaine Laboratories, Inc; 2005. 10. Combi-Screen PLUS® package insert. Analyticon Biotechnologies AG, Lichenfels, Germany.; 2008. 11. DiaScreen© reagent strips [Package insert]. Minneapolis, MN. Hypoguard; 2004. 12. Diuri H-Series® Urinalysis Reagent Strips user's guide. DIRUI Industrial Co., Ltd., Changchun, Jilin Province, China.; 2007. 13. Mission® Urinalysis Reagent Strips package insert. ACON Laboratories, San Diego, CA.; 2007. 14. SELF-STICK® Reagent Strips package insert. ChungDo Pharm. Co., Ltd., Seoul, Korea.; nd. 15. URiSCAN® Urine Strips package insert. YD Diagnostics, Yongin-Si, Korea.; 2007. 16. UriTest 13G® Urine Reagent Strips package insert. Uritest Medical Electronic Co., Ltd. GuiLin, P.R China.; 2006. 17. Uro-dip® 10C Urine Reagent Strips package insert. Erba Diagnostics Mannheim GmbH, Mannheim, Germany.; nd. 18. URS® Urine Reagent Strips for Urinalysis package insert. Teco Diagnostics, Anaheim, CA.; 2005. 19. Hinberg IH, Katz L, Waddell L. Sensitivity of in vitro diagnostic dipstick tests to urinary protein. Clin Biochem 1978;11(2):62–64. 20. James GP, Bee DE, Fuller JB. Proteinuria: accuracy and precision of laboratory diagnosis by dipstick analysis. Clin Chem 1978;24: 1934–1939. 21. Brody LH, SaUaday JR, Armbruster K. Urinalysis and the urinary sediment. Med Clin North Am 1971;55:243–266. 22. Douglas AP, Kerr DNS. A Short Textbook of Kidney Diseases. Great Britain: Pitman Press, 1971. 23. Weller JM. The urinary system. In: Miller SE, Weller JM, eds. Textbook of Clinical Pathology. 8th Ed. Baltimore: Williams & Wilkins Co, 1971: 528–555. 24. Krupp MA, Sweet NJ, Jawetz E, et al. Physician’s Handbook. 19th Ed. Los Altos, CA: Lange Medical Publications, 1979. 25. Brandt R, Guyer KE, Banks WL Jr. Urinary glucose and vitamin C. Am J Clin Pathol 1977;68:592–594. 26. Court JM, Davies HE, Ferguson R. Diastix and ketodiastix: a new semiquantitative test for glucose in urine. Med J Aust 1972;1:525–528. 27. Clinitest© reagent tablets [package insert]. Siemens (formerly Elkhart, IN: Bayer Corporation); 1995. 28. Nahata MC, McLeod DC. Noneffect of oral urinary copper ascorbic acid on reduction glucose test. Diabetes Care 1978;1(1):34–35. 29. Smith D, Young WW. Effect of large-dose ascorbic acid on the twodrop Clinitest determination. Am J Hosp Pharm 1977;34:1347– 1349. 30. Stryer L. Biochemistry. San Francisco: WH Freeman and Co, 1975; Stuber A. Screening tests and chromatography for the detection of inborn errors of metabolism. Clin Chim Acta 1972;36:309–313. 31. Weisberg HF. Water, electrolytes, acid-base, and oxygen. In: Davidson I, Henry JB, eds. Todd-Sanford’s Clinical Diagnosis by Laboratory Methods. 15th Ed. Philadelphia: WB Saunders Co, 1974:772–803. 32. Henry RJ. Clinical Chemistry Principles and Technics. New York: Hoeber Medical Division, Harper & Row, 1974. 33. Mayes PA. Ketone Bodies. Brownsville, TX: Proficiency Testing Service, American Association of Bioanalysts, 1973; Test of the Month No. 16. 34. Acetest© [Package insert]. Elkhart, IN: Siemens (formerly Elkhart, IN: Bayer HealthCare LLC) 2006. 35. Burtis CA, Ashwood ER, eds. Tietz Fundamentals of Clinical Chemistry. 5th Ed. Philadelphia: WB Saunders Co, 2001. 36. Bradley M, Schumann GB, Ward PCJ. Examination of urine. In: Henry JB, ed. Todd-Sanford-Davidson’s Clinical Diagnosis and Management by Laboratory Methods. 16th Ed. Philadelphia: WB Saunders Co, 1979. 37. Berman LB. When the urine is red. J Am Med Assoc 1977;237: 2753–2754. 38. Arnow LE. Introduction to Physiological and Pathological Chemistry. 7th Ed. St Louis: CV Mosby Co, 1966. 39. Bauer JD, Ackerman PG, Toro G. Bray’s Clinical Laboratory Methods. 7th Ed. St Louis: CV Mosby Co, 1968. 40. Sisson JA. Handbook of Clinical Pathology. Philadelphia: JB Lippincott Co, 1976. 41. Smith BC, Peake MJ, Fraser CG. 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Urine and the Urinary Sediment. 2nd Ed. Springfield, IL: Charles C Thomas, 1957. 50. Sobotka H, Luisada-opper AV, Reiner M. A new test for bilirubin in urine. Am J Clin Pathol 1953;23:607–609. 51. Simmons JS, Gentskow CJ. Medical and Public Health Laboratory Methods. Philadelphia: Lea & Febiger, 1955. 52. Hager CB, Free AH. Urine urobilinogen as a component of routine urinalysis. Am J Med Technol 1970;36:227–233. 53. James GP, Paul KL, Fuller JB. Urinary nitrite and urinary-tract infection. Am J Clin Pathol 1978;70:671–678. الدكتور/محمد الختاتنة

37 REFERENCES: Textbook: Graff’s Textbook of Routine Urinalysis and Body Fluids 1. Multistix© 10 SG reagent strips [Color chart]. Tarrytown, NY: Bayer Corporation Diagnostics Division, 1996. 2. Multistix© 10 SG reagent strips [Package insert]. Tarrytown, NY: Siemens Healthcare Diagnostics Inc, 2008. 3. de Wardener HE. The Kidney. 3rd Ed. Boston: Little, Brown & Co, 1967. 4. James GP, Bee DE, Fuller JB. Accuracy and precision of urinary pH determinations using two commercially available dipsticks. Am J. Clin Pathol 1978;70:368–374.


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