meaning that the test should not be taken as conclusive. Negative results can be obtained in the presence of non nitrate-reducing microorganisms. Nitrite-reducing bacteria need to remain in contact with nitrate for long enough to produce detectable amounts (first urine produced in the morning or at least with a urine retention of 4 hours). Large numbers of bacteria can react to reduce nitrite to nitrogen, which will give a false negative result. The use of antibiotics will inhibit bacterial metabolism causing negative results even though bacteria are present. In addition some substances such as ascorbic acid will compete with the Greiss reaction giving unrepresentatively low readings. Leukocytes test It is normal to find up to 3 (occasionally 5) leukocytes per high power field (40X) in a urine sample, with women having slightly higher results owing to vaginal contamination. Higher numbers indicate urinary infection. The urine test strip test for white blood cells detects leukocyte esterase, which is present in azurophilic granules of monocytes and granulocytes (neutrophilic, eosinophilic and basophilic). Bacteria, lymphocytes and epithelial cells from the genitourinary tract do not contain esterases. Neutrophil granulocytes are the leukocytes most commonly associated with urinary infections. A positive test for leukocyte esterase normally indicates the presence of bacteria and a positive nitrite test (although it is not always the case). Infections caused by Trichomonas, Chlamydia and yeasts produce leukocyturia without bacteriuria. The inflammation of the renal tissues (interstitial nephritis) can produce leukocyturia, in particular toxic interstitial nephritis with predominant eosinophils. The test for leukocyte esterase is purely indicative and should not be solely relied on for diagnosis, as it does not replace microscopic or urine culture examinations. The urine test strip reaction is based on the action of leukocyte esterase in catalysing the hydrolysis of an ester of indolecarboxylic acid. The indoxyl that is liberated combines with a diazonium salt in order to produce a violet coloured azole dye. 1) Reaction catalysed by leukocyte esterase Indolecarboxylic acid ester → Indoxyl + Acid 2) In acid medium Indoxyl + Diazonium salt → Violet azole dye The esterase reaction needs about 2 minutes to take place. The presence of strong oxidising agents or formaldehyde can cause false positives. False negative results are associated with elevated concentrations of protein (greater than 500 mg/dL), glucose (greater than 3 g/dL), oxalic acid and ascorbic acid. Urine with a high specific gravity can also cause leukocyte crenation, which can