title: Evaluation of calibrated 1 and 10 microl loops and dipslide as compared to pipettes for detection of low count bacteriuria in vitro.
abstract: Criteria for defining urinary tract infection have changed as the importance of low count bacteriuria in patients with symptoms of infection has been recognized. Little has been published, however, regarding the ability of routine laboratory methods to meet this new challenge. We compared 1 microl and 10 microl calibrated plastic loops (Nunc, Denmark), to a commonly used dipslide method (Uricult, Orion, Finland) using a 0.1 ml pipette as the gold standard. Four typical uropathogens, E. coli, P. aeruginosa, S. saprophyticus and E. faecalis, were mixed in pooled human urine in numbers representing low counts, i.e. 10(2), 10(3), and 10(4), together with 10(5) colony-forming units (cfu)/ml, and sampling with the four methods was performed 20 times for each with each bacterium and each dilution. Colony counts showed that in many cases the 1 microl loop did not deposit any bacteria on the agar plate when colony counts were lower than 10(4) cfu/ml. The 10 microl loop constantly deposited 1.5 times higher numbers of bacteria than predicted, which correlated with the results of weighing the amount of water sampled, i.e. 15.24 (1.55) microg (mean (SD)). The dipslide counts correlated with the 10 microl loop although the agar mounts on the slides absorbed water equal to about 200 microl. There were no significant differences in the counts of the four different bacteria. Variation coefficients increased with decreasing volumes sampled, but the 10 microl loop was found sufficiently exact to detect counts from 10(3) to 10(4), since only 10-fold multiples are usually reported for urine cultures.
authors.affiliation: Department of Clinical Microbiology, Statens Serum Institut, Copenhagen, Denmark. nfm@ssi.dk
authors.forename: N
authors.initials: N
authors.lastname: Frimodt-Møller
authors.forename: F
authors.initials: F
authors.lastname: Espersen
mesh_terms: D001431:Bacteriological Techniques / Q000295:instrumentation*; D001437:Bacteriuria / Q000175:diagnosis / Q000382:microbiology*; D002138:Calibration; D015169:Colony Count, Microbial / Q000379:methods; D013293:Enterococcus faecalis / Q000302:isolation & purification; D004926:Escherichia coli / Q000302:isolation & purification; D006801:Humans; D011550:Pseudomonas aeruginosa / Q000302:isolation & purification; D013210:Staphylococcus / Q000302:isolation & purification