MULTIPLE REAGENT STRIP FOR CHEMICAL EXAMINATION OF URINE

Two Major Types of Reagent Strips:

1. Multistix (Siemens Medical Solution Diagnostics)
2. Chemstrip (Roche Diagnostics)
Formed elements: sink to the bottom, will be undetected if not properly
mixed

Good light source: for accurate interpretation of color reactions

Distilled Water: not recommended as (-) control as strips are designed to
perform at ionic concentration similar to urine
Phenazopyridine compounds: orange pigments causing masking of
reactions
Non reagent strip testing procedures: tablets and liquid chemicals

PARAMETE
R
GLUCOSE

PRINCIPLE
Double sequential
enzyme reaction

REAGENT
(COLOR REACTION)
Glucose oxidase
Peroxidase

5.5 mmol/L
M: potassium iodide
(green-brown)
C: Tetramethylbenzidine
(yellow-brown)

BILIRUBIN
100 mg/dL

KETONE
No ketones
are present
in normal
urine
SPECIFIC
GRAVITY
Random:
1.001-1.035
24 hr:
1.016-1.022
BLOOD
Green spots
w/in 60 secs
recommend
investigation

Coupling of
bilirubin with
diazotized
dichloroaniline salt
in a very acetic
medium
Development of
colors from buffpink (-) to purple
(acetoacetic acid
reacts w/
nitroprusside)
pKa change of
certain pretreated
polyelectrolytes in
relation to ionic
concentration

Psuedoperoxidase
activity of
hemoglobin to
catalyze between
H2O2 &
tetramethylbenzidi
ne

M: 2,4-dichloroaniline diazonium
salt
C: 2,6-dichlorobenzenediazonium salt
(tan to pink to violet)
Sodium nitroprusside
Glycine (C)
(yellow to purple)

M: Poly Bromthymol blue

LIMITATION
FALSE (+)
FALSE (-)
Contamination of
ascorbic acid
oxidizing agent &
ketones
detergent
specific gravity
temperature
Improperly
preserved
specimens

C:dimethyldihydroperoxyhexane
tetramethylbenzidine
(yellow to green-blue)

CLINICAL SIGNIFICANCE

Glucose Oxidase
Moore-Hellers
Test
Copper Reduction
Test
a. Clintest
Other Sugar
reducing Tests:
a. Osazone test
b. Fehlings test
c. Nylanders
test
Icotest tablets
Foam Test

HYPERGLYCEMIA RELATED
DM, pancreatitis, pancreatic CA,
acromegaly, cushing syndrome,
hyperthyroidism,
pheohromocytoma, CNS damage,
stress, gestational diabetes

Improperly
preserved
specimen

Acetest tablets

Diabetic acidosis, insulin dosage

monitoring, starvation,
malabsorption, pancreatic
disorders, strenuous exercise,
vomiting, inborn errors of amino
acid metabolism
Hydration status
Loss of renal tubular concentrating
ability
Diabetes insipidus
Determination of unsatisfactory
specimens due to low conc

Indican (indoxyl
sulfate)
Metabolites of
Lodine (etodolac)
Highly pigmented
specimen
Phenazopyridine
Highly pigmented
specimen
Levodopa
Mesna
Medications w/ free
sulfhydryl groups
High protein
concentration

Exposure to light
Ascorbic acid
>25mg/dL
High nitrite conc.

Highly alkaline
urine (>6.5)

Urinometery
Refractometery
Harmonic
Oscillation
Densitometry

Strong oxidizing
agents
Bacterial
peroxidises
Menstrual
contamination

High specific
gravity/ crenated
cells
Formalin
Captopril
High nitrite conc.
Ascorbic acid
>25mg/dL
Unmixed specimen

Hemoglobinuria vs
Myoglobinuria
Plasma
comparison
Immunoassay for
myoglobin levels

C:ethyleneglycoldiaminoethyleth
ertetraacetic acid
Bromthymol blue
(blue [1.000 alkaline] to yellow
[1.030 acid])
M: Diisopropylbenzene
dehydroperoxide 3,31,5,51tetramethylbenzidine

OTHER TESTS

RENAL ASSOCIATED
Fanconi syndrome, advance renal
disease, osteomalcia, pregnancy
Hepatitis, cirrhosis, other liver
disorders, biliary obstruction

HEMATURIA (renal calculi,

glomerulonephritis, pyelonephritis,
tumor, trauma, toxic chemicals,
anticoagulants, strenuous exercise)
HEMOGLOBINURIA (transfusion
reaction, hemolytic anemia, severe
burns, infection, malaria, strenuous
exercise, blood cell trauma, brown
recluse spider bites)
MYOGLOBNURIA (muscular trauma,
crush syndromes, prolonged coma,
convulsions, muscle-wasting
disease, alcoholism, drug abuse,

pH
5-9

PROTEIN

Protein
presence in
the urine is
not a normal
occurrence

Double indicator
system of methyl
red and
bromthymol blue

Both M &C measures pH=5-9

Protein error of
indicators
principle:
Yellowish (-)
Greenish-blue (+)

Buffer: acid (pH=3)

Methyl red (pH=4-6)

(red-yellow)
Bromthymol blue (pH=6-9)
(yellow-blue)

M: Tetrabromphenol blue
C: 3,3,5,5-tetrachloropheno3,4,5,6tetrabromosulfonphthalein
(Yellow to blue-green)

Runover between
pH and protein
testing area

Runover
Excess urine
Inproper procedure

Lithmus paper/
Nitrazine paper (if
pH urine is the
only test needed
to be done)

Highly buffered
alkaline urine
Pigmented
specimen
Phenazopyridine

Proteins other that

albumin

Foam Test
Sulfosalicylic Acid
Precipitation Test

microalbuminuria
MICROALBUMINUR
IA TESTS:
Immunologic
Assays
a. Micral Test
b. Immunodip

Quaternary
ammonium
compounds
(detergents)

UROBILINO
GEN
3.2-16
umol/L
0.2-1.0
mg/dL
NITRITE
No nitrite
are present
in normal
urine
abnormal
conc: 4080%

Mod. Ehrlichs
reaction

M: p-diethylaminobenzaldehyde
C: 4-methoxybenzen-diazoniumtetrafluoroborate
(shades of pink)

1. Nitrate nitrite
by gram (-)
bacteria
2. Greiss reaction
(nitrite at acidic pH
reacts with
aromatic amine to
form diazonium
compound that
reacts with
Hydroxytetra

M: p-arsinilic acid
Tetrahydrobenzo(h)quinolin-3-ol
C: Sulfanilamide
Hydroxytetra
hydrobenzoquinoline
(white to pink)

RENAL (Glomerular disorders, IC

disorders, Amyloidosis)
TUBULAR DISORDERS (Fanconi,
toxic agents, heavy metals, severe
viral infections)
POSTRENAL (lower UTI,
injury/trauma, menstrual
contamination, sperm, vaginal
secretion)

Antiseptics
Chlorhexidine
High specific
gravity
Loss of buffer due
to prolong
exposure of strip to
the specimen
Porphobilinogen
Indican
p-aminosalicylic
acid
sulphonamides
methyldopa
chlorpromazine
Improperly
preserved
specimen
Highly pigmented
urine

extensive exertion, cholesterol

lowering statin meds)
Respiratory/ metabolic
acidosis/alkalosis
Defects in renal tubular secretion
& reabsorption
Renal calculi formation
Treatment of UTI
Precipitation/ identification of
crystals
Determination of unsatisfactory
specimen
PRERENAL (IV hemolysis, muscle
injury, APR, MM)

Toxic agents, diabetic nephropathy,

strenuous exercise, dehydration,
hypertension, pre-eclampsia,
orthostatic/ postural proteinuria
Old specimen
Formalin
preservation
High nitrite conc

Ehrlich Tube Test

Watson-Schwartz
Differentiation Test
Hoesch Screening
Test for
Porphobilinogen

Early detectionof liver disease

Liver disease, hepatitis, cirrhosis,
CA
Hemolytic disease

Non reductase
bacteria
Insufficient contact
time
Lack of urinary
nitrate
Large quantity of
bacteria
Antibiotics
Ascorbic acid
>25mg/dL

Nitrite Test

Cystitis
Pyelonephritis
Evaluation of antibiotic therapy
Monitoring of patients at high risk
for UTI
Screening urine culture specimen

LEUKOCYT
E
ESTERASE
Lekocytes
not normally
found in
urine

hydrobenzoquinoli
ne
Hydrolysis of
derivatized pyrrole
amino acid ester to
liberate 3hydroxyl-5-phenyl
pyrrole

M: Derivatized pyrrole amino acid

ester
Diazonium salt
C: Indoxylcarbonic acid ester
Diazonium salt
( lavender to purple)

Strong oxidizing
agents
Formalin
Highly pigmented
urine
Nitrofurantoin

High specific
gravity
High conc. of
CHON, glucose,
oxalic acid,
ascorbic acid,
gentamicin,
cephalosporin,
tetracycline,
inaccurate timing

Leukocyte
Esterase Test
Evaluation of pH,
protein, nitrite

UTI
Urinary tract inflammation
Screening of urine culture
specimen