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URINALYSIS (MLT 305) LECTURE TWO. Dr. Essam H. Jiffri. BLOOD. - Blood may be present in the urine either in the form of intact red blood cells (hematuria) or as the red blood cell destruction product hemoglobin (hemoglobinuria).

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blood
BLOOD
  • - Blood may be present in the urine either in the form of intact red blood cells (hematuria) or as the red blood cell destruction product hemoglobin (hemoglobinuria).
  • - Blood present in large quantities can be detected visually; hematuria produces a cloudy red urine and hemoglobinuria appears as a clear red specimen.
blood3
BLOOD
  • Any amount of blood greater than 5 cells per microliter of urine is considered clinically significant, it is not possible to rely on visual examination to detect the presence of blood.
  • - The microscopic examination can be used to differentiate between hematuria and hemoglobinuria.
blood4
BLOOD
  • REAGENT STRIP REACTIONS
  • - Chemical tests for blood utilize the pseudoperoxidase activity of hemoglobin to catalyze a reaction between:
  • - hydrogen peroxide and the chromogen tetramethylbenzidine
  • to produce an oxidized chromogen, which has a green-blue color.
  • hemoglobin
  • H2O2 + Chromogen Oxidized Chromogen + H2O
  • peroxidase
blood5
BLOOD
  • REAGENT STRIP REACTIONS
  • - Two color charts are provided that correspond to the reactions that occur with hemoglobinuria and hematuria.
  • - In the presence of free hemoglobin:
  • - A strongly positive blue will appear on the pad.
  • - In contrast, intact red blood cells:
  • - A speckled pattern will appear on the pad.
blood6
BLOOD
  • CLINICAL SIGNIFICANCE OF URINE BLOOD
  • Hematuria
  • 1. Renal calculi
  • 2. Glomerulonephritis
  • 3. Pyelonephritis
  • 4. Tumors
  • 5. Trauma
  • 6. Exposure to toxic chemicals or drugs
  • 7. Strenuous exercise
blood7
BLOOD
  • CLINICAL SIGNIFICANCE OF URINE BLOOD
  • Hemoglobinuria
  • 1.Transfusion reactions
  • 2.Hemolytic anemia
  • 3. Severe burns
  • 4. Infections
  • 5. Strenuous exercise
blood8
BLOOD
  • CLINICAL SIGNIFICANCE OF URINE BLOOD
  • Myoglobinuria
  • 1. Muscular trauma
  • 2. Prolonged coma
  • 3. Convulsions
  • 4. Muscle-wasting diseases
  • 5. Extensive exertion
bilirubin
BILIRUBIN
  • - The appearance of bilirubin in the urine is the first indication of liver disease and is often detected long before the development of jaundice.
  • - Bilirubin provides early detection of hepatitis, cirrhosis, gallbladder disease, and cancer, and should be included in every routine urinalysis.
bilirubin bilirubin a highly pigmented yellow compound is a degradation product of haemoglobin
BilirubinBilirubin, a highly pigmented yellow compound, is a degradation product of haemoglobin
bilirubin11
Bilirubin
  • OXIDATION TESTS (FOUCHET’S TEST)
  • - Urine containing bilirubin usually appears dark yellow or amber and produces a yellow foam when shaken, this foam test was actually the first test for bilirubin.
  • - Oxidation tests utilize the ability of feric chloride dissolved in trichloracetic acid (Fouchet's reagent) to oxidize bilirubin to biliverdin, producing a green color.
bilirubin12
Bilirubin
  • REAGENT STRIP (DIAZO) REACTIONS
  • Routine testing for urinary bilirubin by reagent strip utilizes the diazo reaction, in an acid medium to produce colors ranging from increasing degrees of tan or pink to violet, respectively.
  • Questionable results should be retested using the Ictotest which produces a more sharply colored diazo reaction.
  • - Colors other than blue or purple appearing on the mat are considered negative.
bilirubin13
Bilirubin
  • CLINICAL SIGNIFICANCE OF URINE BILIRUBIN
  • 1. Hepatitis
  • 2. Cirrhosis
  • Biliary obstruction
bilirubin14
- The presence or absence of bilirubin can be used in determining the cause of clinical jaundice.

( Urine Bilirubin and Urobilinogen in Jaundice )

Bilirubin
urobilinogen
UROBILINOGEN
  • Like bilirubin, urobilinogen is a bile pigment that results from the degradation of hemoglobin.
  • It is produced in the intestine by the oxidation of bilirubin by the intestinal bacteria.
  • - Approximately half of the urobilinogen is reabsorbed from the intestine into the blood, recirculates to the liver, and is secreted back into the intestine through the bile duct.
urobilinogen16
UROBILINOGEN
  • The urobilinogen remaining in the intestine is excreted in the feces, where it is oxidized to urobilin, the pigment responsible for the characteristic brown color of the feces.
  • Urobilinogen appears in the urine because, as it circulates in the blood en route to the liver, it passes through the kidney and is filtered by the glomerulus.
  • - A small amount of urobilinogen less than 1 mg/dl or 1 Ehrlich unit is normally found in the urine.
urobilinogen18
UROBILINOGEN
  • EHRLICH'S TUBE TEST
  • The reagent used in all tests was
  • p-dimethylaminobenzaldehyde (Ehrlich's reagent), addition of Ehrlich's reagent to urine containing urobilinogen produces a cherry red color.
  • - Positive results in dilutions greater than 1 to 20 were considered significant.
urobilinogen19
UROBILINOGEN
  • EHRLICH'S TUBE TEST
  • An Ehrlich unit is essentially equal to 1 mg of urobilinogen.
  • - Normal values for females are 0.1 to 1.1 Ehrlich units; and for males, 0.3 to 2.1 Ehrlich units, these values are based on the recommended 2-hour specimen collected after the noon meal between 2 and 4 PM,which is the time of greatest urobilinogen excretion.
urobilinogen20
UROBILINOGEN
  • CLINICAL SIGNIFICANCE OF URINE UROBILINOGEN
  • 1. Early detection of liver disease
  • 2. Hemolytic disorders
nitrite
NITRITE
  • - The reagent strip test for nitrite provides a rapid screening test for the presence of urinary tract infection.
nitrite22
NITRITE
  • REAGENT STRIP REACTIONS
  • Nitrite is detected by the Greiss reaction:
  • - nitrite at an acidic pH reacts with an aromatic amine to form a diazonium compound that react with quinolin compound to produce a pink color.
nitrite23
NITRITE
  • CLINICAL SIGNIFICANCE OF URNE NITRITE
  • 1. Cystitis (initial bladder infection)
  • 2. Pyelonephritis (an inflammatory process of the kidney and adjacent renal pelvis)
  • 3.Evaluation of antibiotic therapy
  • 4. Monitoring of patients at high risk for urinary tract infection
  • 5. Screening of urine culture specimens (detection of bacteruria)
spicific graviiy
SPICIFIC GRAVIIY
  • The addition of a specific gravity testing area to Multistix has eliminated:
  • time-consuming step in routine urinalysis
  • provided a convenient method for routine screening.
  • - It is not recommended to replace osmometry or refractometry for critical fluid monitoring.
spicific graviiy25
SPICIFIC GRAVIIY
  • REAGENT STRIP REACTION
  • - The test is based on the change in pK- (dissociation constant) of poly methyl vinyl- ethermaleic anhydride.
spicific graviiy26
SPICIFIC GRAVIIY
  • CLINICAL SIGNIFICANCE OF URINE SPECIFIC CRAVITY
  • 1. Patient hydration and dehydration
  • 2. Loss of renal tubular concentrating ability
  • 3. Diabetes insipidus
  • 4. Determination of unsatisfactory specimens due to low concentration
lukocytes
LUKOCYTES
  • - One of the most frequent findings in the routine urinalysis is the presence of leukocytes, indicating a possible infection of the urinary tract.
  • - Detection of leukocytes was previously made only by microscopic examination of the urinary sediment.
lukocytes28
LUKOCYTES
  • REAGENT STRIP REACTION
  • The chemical reaction is enzymatic, utilizing esterases present in granulocytic white blood cells to hydrolyze indoxylcarbonic acid ester to produce indoxyl, which reacts with a diazonium salt to create a purple color.
  • leukocyte
  • indoxylcarbonic acid ester indoxyl + diazonium salt (purple color)
  • esterases
  • - It is recommended that trace reactions be repeated on a fresh specimen and that microscopic examinations be performed on all positive specimens.
lukocytes29
LUKOCYTES
  • CLINICAL SIGNIFICANCE OF URINE LEUKOCYTES
  • 1. Urinary tract infection
  • 2. Screening of urine culture specimens