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The Journal of Medical Microbiology, Vol 45, Issue 1 76-78, Copyright © 1996 by Society for General Microbiology

JOURNAL ARTICLE

A rapid immunoassay method for the direct detection of PCR products: application to detection of TEM beta-lactamase genes

R. Curran, D. C. Talbot and K. J. Towner
Department of Microbiology & PHLS Laboratory, University Hospital, Queen's Medical Centre, Nottingham, UK.

A rapid immunoassay for the detection of specific PCR products is described in which a positive PCR amplification result is detected, usually in less than 5 min, by applying a few drops of the diluted PCR end-product to a small immunoassay sample device. The method was evaluated in comparison with conventional susceptibility tests and isoelectric focusing (IEF) for the detection of TEM-family beta-lactamase genes in 477 Escherichia coli isolates from urine samples. Of 187 isolates identified as presumptive TEM beta-lactamase producers by conventional methods, 185 generated a positive signal in the PCR immunoassay system. Two further signal-positive isolates were recognised when the PCR was repeated. In addition, one of the 276 ampicillin-susceptible isolates gave a positive signal in repeated PCR-immunoassay experiments despite being ampicillin susceptible and failing to give a TEM-type enzyme band in iso-electric focusing experiments.


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