Prepared by:

PRINCESS ALEN I. AGUILAR,RMT

 1. SPECIFIC GRAVITY
PRINCIPLE
M: Poly(methylvinyl ether) maleic anhydride, BTB
REAGENTS
C: Ethyleneglycol-bis (aminoethylether), BTB
COLOR & READING TIME

SOURCES OF F(+): High concentration of protein

INTERFERENCE F(–): Highly alkaline urine

1. Monitoring patient’s hydration.

2. Detection of loss of renal concentrating ability (in cases
of CGN, RF, ATN)
CLINICAL SIGNIFICANCE
3. Diagnosis of diabetes insipidus (Hyposthenuria)
4. Determination of unsatisfactory specimens due to low
concentration
 2. pH
PRINCIPLE
REAGENTS
COLOR & READING TIME
SOURCES OF INTERFERENCE Runover from adjacent pads Old specimens
1. Respiratory or metabolic acidosis/ alkalosis 2
2. Renal tubular acidosis
3. Renal calculi formation
CLINICAL SIGNIFICANCE
4. Treatment of UTI
5. Precipitation/ identification of crystals
6. Determination of unsatisfactory specimen
Important in the identification of crystals and
ADDITIONAL NOTES
determination of unsatisfatory specimens
 pH
• Normal pH:
– Random  4.5-8.0
– 1st Morning  5.0-6.0
• pH of 9 = _________________________
• ______________occurs after meals due to withdrawal of
hydrogen ions for the purpose of secretion of HCl
• Causes of acid urine: emphysema, diabetes mellitus,
starvation, diarrhea, dehydration, acid producing bacteria,
high protein diet, medications , _________________ (tx
for UTI)
• Causes of alkaline urine: hyperventilation, vomiting,
______________________, urease-producing bacteria,
vegetarian diet, old specimens , __________________
 3. PROTEIN
_______________________________
PRINCIPLE
indicator is sensitive only to ALBUMIN
M: Tetrabromphenol blue
REAGENTS
C: Tetrachlorophenol tetrabromosulfonphthalein
COLOR & READING TIME
F(+): Highly buffered alkaline urine, pigmented
specimens, chlorhexidine, phenazopyridine, QACs
SOURCES OF INTERFERENCE (detergents), antiseptics, loss of buffer, high SG F(–
):Proteins other than albumin, high salt concentration,
microalbuminuria
NORMAL:
____________________________________________
Degrees of proteinuria:
CLINICAL SIGNIFICANCE
a. Mild – < 1.0 g/day
b. Moderate – 1.0-4.0 g/day
c. Heavy – > 4.0 g/day
MOST INDICATIVE OF RENAL DISEASE
ADDITIONAL COMMENT WHITE FOAM IN URINE
 PROTEIN
1. __________________  Major serum protein
found in the urine
Normal values:
<10 mg/dL or 100mg/24 hrs (Strasinger)
<150mg/dL (Henry)
2. OTHER PROTEINS
a. Serum & Tubular microglobulins
b. Tamm-Horsfall protein (aka Uromodulin)
c. Protein derived from prostatic and vaginal
secretion
 Types of proteinuria:
A. Pre-renal – caused by conditions that affect
plasma prior to its reaching the kidney
 intravascular hemolysis
 muscle injury
 severe infection and inflammation;
 ____________________- proliferation of Igs-
producing plasma cells (BJP)
 Test  SPE, Immunofixation electrophoresis
 Urine = precipitates at 40-600C (cloudy) and
dissolves at 1000C
Classification of proteinuria
Classification of proteinuria
 Types of proteinuria:
B. Renal
A. Glomerular Proteinuria
1. Diabetic nephropathy- decreased GFR, may lead to renal failure
» Indicator: microalbuminuria
2. _________________________________________
 Proteinuria when standing due to increased pressure to renal veins

ORTHOSTATIC PROTEINURIA CLINICAL PROTEINURIA

First Morning
2 hours after standing
3. Amyloidosis, Glomerulonephritis, Autoimmune Disorders, Toxic
Agents, Hypertension, Strenuous Exercise, Preeclampsia,
Dehydration,
B. Tubular Proteinuria– Reabsorption defective
 Fanconi syndrome,
 toxic agents
 severe viral infections
 Types of proteinuria:
C. Post-renal – after
1. lower UTI;
2. injury or trauma;
3. menstrual contamination;
4. prostatic fluid;
5. spermatozoa;
6. vaginal secretions
a) Heat and Acetic Acid Test
– Grading:
• diffused cloud (+1);
• granular cloud (+2);
• distinct flocculi (+3);
• large flocculi (+4) TESTS
b) SSA Test/ Cold Protein Precipitation FOR
PROTEIN
b) SSA Test/ Cold Protein Precipitation
 Reacts equally to all forms of protein
 RGT: 3mL of 3% SSA + 3mL centrifuged urine 
_______________________________
P
• False (+): mucin, uric acid, penicillin, tolbutamides,
radiocontrast media, sulfonamides, cephalosporins
T R
• False (-): highly buffered alkaline urine
• Correlate with reagent strip results
E F O
c)Tests for microalbuminuria S O T
 Micral test and Immunodip  strip emplying Ab-
Enzyme conjugate that binds albumin T R E
 (-) WHITE ; (+) RED
 Significant values reported as AER S I
 Normal AER = ________________
 Microalbuminuria = _____>300mg N
___________________
 Clinical albuminuria = >200 ug/min
 4. GLUCOSE
PRINCIPLE Glucose oxidase reaction / Double sequential enzyme rxn
M: Glucose oxidase, peroxidase, KI
REAGENTS
C: Glucose oxidase, peroxidase, TMB
COLOR & READING TIME M: greenbrown C: yellowgreen (30 s)
F(+): Oxidizing agents, detergents
SOURCES OF INTERFERENCE F(–): Ascorbic acid, ketones, high SG, low temperatures,
improperly preserved specimens
Types of Glucosuria:
a. Hyperglycemia-associated – diabetes mellitus,
endocrine disorders,( Cushing’s, Pheochromocytoma,
Acromegaly, Hyperthyroidism) pancreatic disorders,
CLINICAL SIGNIFICANCE
CNS disorders, disturbance in metabolism, liver
disease, drugs, gestational diabetes mellitus
b. Renal-associated – renal tubular dysfunction, tubular
necrosis, Fanconi syndrome, osteomalacia, pregnancy
ADDITIONAL COMMENT Most frequently tested in urine; threshold substance
 Benedict’s test
• (Copper Reduction) • _____________________
CuSO4  (+) Cu2O • Procedure: 5 gtts urine + 10 gtts
(Blue) reducing subs H2O + clinitest tablet
 Positive result: _____________ • Rgts:
– ______________ (main reacting rgt)
– NaOH, sodium citrate (for heat
production)
– NaCO3 –elliminates interfering O2
• False(+): other reducing sugars,
ascorbic acid, drug metabolites,
cephalosporin
• False (-):
– oxidizing agents (detergents),
– pass-through phenomenon
• Occurs when >2g/dL sugar is present
• To prevent, use 2gtts urine
• Correlate with reagent strip results
 NOTE:
 Non-specific test for reducing sugars
• Fructose (Levulose)
• Galactose
• Lactose (Glu-Gal)
• Pentose (Xylulose, Arabinose)
• Sucrose (Glu-Fru) – non reducing sugar,
thus negative
 5. KETONE
____________________________________
PRINCIPLE Acetoacetic acid + Na Nitroprusside  purple (+)
(Acetone) (Glycine)
M: Sodium nitroprusside
REAGENTS
C: Sodium nitroprusside and glycine (can detect acetone)
COLOR & READING TIME
F(+): Phthalein dyes, highly pigmented red urine,
SOURCES OF INTERFERENCE levodopa, medications containing SH group
F(–): Improperly preserved specimens
Ketone bodies: Acetone – 2% Acetoacetic acid – 20%
β-hydroxybutyric acid – 78%
Causes of Ketonuria: diabetes mellitus, starvation,
CLINICAL SIGNIFICANCE
fasting, weight reduction, strenuous exercise,
malabsorption, pancreatic disorders, inborn errors of
amino acid metabolism
CONFIRMATORY TEST
• CONTAINS:
– Sodium nitroprusside
– Disodium phosphate 
– Lactose 
• 10 times sensitive to diacetic acid than acetone
• Can detect in urine:
– 5–10 mg/dL of diacetic acid and 20–25 mg/dL of
acetone
1. Place the tablet on a piece of clean, dry white paper.
2. Put one drop of urine, serum, plasma, or whole blood
directly on top of the tablet.
3. For urine, compare the color of the tablet with the color
chart at 30 seconds. For serum or plasma, compare the color
after 2 minutes. For whole blood, remove the clotted blood
from the tablet after 10 minutes and compare the color of
the tablet with the chart.
Note: for serum, plasma and whole blood, the loest limit of
detection s 10mg of diacetic acid per 100mL
1. Gerhart’s, Lindeman’s – diacetic acid
2. Frommer’s, Walhauster’s, Lange’s,
JacksonTaylor’s, Rantzmann’s, Lieben’s –
acetone
3. Legal’s, Rothera’s, Acetest, Ketostix – diacetic
acid and acetone
4. Osterberg, Hart’s test – β-hydroxybutyric acid
 6. BLOOD
Hgb
PRINCIPLE
H2O2 + Chromogen  oxidized chromogen + H2O
Pseudoperoxidase
M:Diisopropylbenzene dihydroperoxide TMB
REAGENTS
C: 2,5-dimethyl 2,5dihydroperoxyhexane TMB
(-) yellow ; (+)Blue-green (60 s)
COLOR & READING TIME Uniformed green/blue color= Hgb/Mgb
Speckled/Spotted = Hematuria (intact RBCs)
F(+): Strong oxidizing agents, bacterial peroxidases, menstrual
contamination
SOURCES OF INTERFERENCE
F(–): High SG, crenated cells, formalin, captopril, nitrite, ascorbic
acid, unmixed specimen
a. Hematuria
CLINICAL SIGNIFICANCE b. Hemoglobinuria
c. Myoglobinuria
CONFIRMATORY TEST
 HEMATURIA HEMOGLOBINURIA MYOGLOBINURIA

Seen in: Seen in: Seen in: Rhabdomyolysis

 Glomerulonephritis  Intravascular hemolysis Heme portion of the
 Renal calculi  Transfusion rxns myoglobin is toxic to the
Microscopic: INTACT RBCS  Hemolytic Anemia renal tubules
 OTHER TESTS FOR URINE BLOOD
• Enzyme assays
• Absorption Spectrophotometry
• Immunodiffusion Technique
• Electrophoresis
 7. BILIRUBIN
___________________________
PRINCIPLE
B2 + Diazonium salt  Azodye
M: 2,4-dichloroaniline diazonium salt
REAGENTS
C:2,6-dichlorobenzene diazoniumtetrafluoroborate
COLOR & READING TIME Tan, pink, or violet (30 s)
F(+): Highly pigmented urine, indican, phenazopyridine, lorpromazine
SOURCES OF INTERFERENCE (Thorazine), metabolites of Lodine , chF(–): Specimen exposure to light,
ascorbic acid >25 mg/dL, high concentration of nitrite
1. Diagnosis of hepatitis, cirrhosis, other liver disorders, and biliary
obstruction
CLINICAL SIGNIFICANCE
2. Determination is more significant when combined with serum
bilirubin and urine urobilinogen
CONFIRMATORY TEST
 Conjugated bilirubin (water soluble)
ADDITIONAL COMMENTS  Early indication of liver disease
 Amber urine with yellow foam
 able to detect as little as
0.05 mg/dL bilirubin
Contains:
 P-nitrobenzene-diazonium
p-toluenesulfonate
 SSA (provide acidity and act
with Sodium carbonate to
provide effervescence
 Sodium carbonate
 Boric acid
Procedure:
1. Place five drops of urine on
one square of the special test
mat supplied with Ictotest.
2. Place a tablet in the center of
the moistened area.
3. Flow two drops of water onto
the tablet so that the water
runs off of the tablet and onto
the mat.
4. Observe the color of the mat
around the tablet at the end
of 30 seconds. If a blue or
purple color develops, the test
is positive.
 Other Tests For Bilirubin
• Foam Shake Test
• Oxidation Test (Gmelin or Fouchet’s method)
 Acidic oxidation of bilirubin into a rainbow
array of colors:
– green (biliverdin)
– blue (bilicyanin)
– yellow (choletelin)
 8. UROBILINOGEN
PRINCIPLE ____________________________: UBG +PDAB  (+) RED
M: paradiethylaminobenzaldehyde
REAGENTS
C: 4-methoxybenzene diazonium tetrafluoroborate
COLOR & READING TIME
F(+): PBG, indican, procaine, p-aminosalicylic acid,
SOURCES OF INTERFERENCE sulfonamides, methyldopa, chlorpromazine, pigmented urine
F(–): Old specimens, formalin, high concentration of nitrite
1. Only a small amount is normally found in urine.
2. Useful for the early detection of liver disease and for the
CLINICAL SIGNIFICANCE diagnosis of hemolytic disorders, hepatitis, cirrhosis, and
carcinoma
3. Absence in urine may indicate biliary obstruction
TEST USED TO DIFFERENTIATE
UROBILINOGEN TO
PORPHOBILINOGEN
 (< 1.0 mg/dL or <1.0 Ehrlich unit)
ADDITIONAL COMMENTS
 Specimen: Afternoon urine (2-4PM due to alkaline tide)
 1. Ehrlich’s Tube Test
• Reagent: p-dimethylaminobenzaldehyde
• (+) Result: cherry red color
2. WATSON-SCHWARTZ TEST
• Uses extraction with organic solvents _____________& ______________
• Sodium acetate  enhances reaction
• INVERSE EHRLICH REACTION
• Rapid screening test for _________________
(>2 mg/dL)
Procedure:
2 gtts urine + 2mL Hoesch rgt
( Ehrlich’s rgt in 6M or 6N HCl)
 9. NITRITE
PRINCIPLE
M: p-arsanilic acid, tetrahydrobenzoquinolinol
REAGENTS C: sulfanilamide, 3hydroxy- 1,2,3,4- tetrahydro- 7,8-
benzoquinoline
Uniform Pink (+) = 100,000 org/mL
COLOR & READING TIME
not pink spots/edge (-) (60 s)
F(+): Old specimen, highly pigmented urine
F(–):Non-reductase-containing bacteria, lack of urinary nitrate,
SOURCES OF INTERFERENCE insufficient contact time between bacteria and nitrate,
bacteria converting nitrite to nitrogen, antibiotics, ascorbic
acid, high SG
1. Diagnosis of cystitis and pyelonephritis
2. evaluation of antibiotic therapy
CLINICAL SIGNIFICANCE
3. Monitoring of patients at high risk for UTI
4. Screening of urine culture specimens
 _______________________________________________
ADDITIONAL COMMENTS
 Specimen: 1st morning or 4hr urine
 10. LEUKOCYTE ESTERASE
PRINCIPLE
M:Derivatized pyerole amino acid ester, diazonium salt
REAGENTS
C: Indoxylcarbonic acid ester, diazonium salt
____________________________ (+) WBC (exc. Lymphocyte),
COLOR & READING TIME
histiocyte, Trichomonas vaginalis = contain esterase
F(+): Strong oxidizing agents, formalin, highly pigmented urine,
nitrofurantoin
SOURCES OF INTERFERENCE
F(–): protein, glucose, oxalic acid, ascorbic acid, gentamicin,
cephalosporins, tetracyclines
1. Detection of bacterial and nonbacterial UTI
CLINICAL SIGNIFICANCE 2. Inflammation of the urinary tract
3. Screening of urine culture specimens
 Strip can detect even lysed WBCs
ADDITIONAL COMMENTS
 List of References
 Lillian Mundt & Kristy Shanahan, Graff’s Textbook
of Urinalysis and Body Fluids, 2nd Ed.
 Susan Strassinger & Marjorie Di Lorenzo,
Urinalysis and Body Fluids, 5th & 6th Ed.
 Erol Coderres,RMT-AUBF notes
 Roderick Balce, RMT-CEU Professor AUBF Notes
 Rexson R. Julian ,RMT- PPT notes
 Ridley,J. et al, Essentials of Clinical Laboratory
Science, ©2011