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OF URINE
CHAPTER 5
• Interference
– No known substances interfere with urinary pH measurements performed by
reagent strips
• Traditional principle
– Protein error of indicators
– Certain indicators change color in the presence of
protein at a constant pH
– Protein accepts H+ from the indicator, increased
sensitivity to albumin due to more amino groups to
accept H+ than other proteins
pH 3.0
Indicator (H+) + Protein → Protein + H+
(Yellow) → Indicator – H+
(Green/Blue)
Cu(CRE) peroxidase
Glucose oxidase
Glucose + O2 (air) → gluconic acid + H2O2
Peroxidase
H2O2 + chromogen → oxidized colored chromogen + H2O
• Chromogens used
– Potassium iodide (green to brown) (Multistix)
– Tetramethylbenzidine (yellow to green) (Chemstrip)
• Reporting results
– Neg, trace, 1+, 2+, 3+, 4+
– 100 mg/dL to 2 g/dL
– 0.1% to 2%
Heat
CuSO4 (cupric sulfide) + reducing substance -----
Alkali
Acid
para-arsanilic acid or sulfanilamide + NO2 —————→ diazonium salt
(nitrite)
Acid
diazonium salt + tetrahydrobenzoquinolin —————→ pink azodye
• False-positive
– Old specimens (bacterial multiplication)
– Highly pigmented urine
– Pink edge or spotting on reagent strip is considered
negative
– Check automated readers manually for color
interference
Leukocyte
indoxylcarbonic acid ester —————→ indoxyl + acid indoxyl
Esterase
Acid
+ diazonium salt ————→ purple azodye
Copyright © 2014. F.A. Davis Company
Reagent Strip Reactions (cont’d)
• LE reaction requires the longest time of all the reagent
strip reactions
– 2 minutes
• Reported as
– Negative
– Trace
– Small: 1+
– Moderate: 2+
– Large: 3+
• Trace readings may not be significant and should be
repeated on a fresh specimen
Copyright © 2014. F.A. Davis Company
Reaction Interference
• False-positive
– Strong oxidizing agents
– Formalin
– Highly pigmented urine, nitrofurantoin
• False-negative
– High concentrations of protein, glucose, oxalic acid, ascorbic
acid
– Crenation from high specific gravity
– Inaccurate timing: must have 2 min
– Presence of the antibiotics; gentamicin, cephalosporins,
tetracyclines
Copyright © 2014. F.A. Davis Company
Specific Gravity