A rapid immunoassay method for the direct detection of PCR products: application to detection of TEM beta-lactamase genes.
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.
Curran
R
R
Department of Microbiology & PHLS Laboratory, University Hospital, Queen's Medical Centre, Nottingham, UK.
Talbot
D C
DC
Towner
K J
KJ
D001437:Bacteriuria / Q000382:microbiology; D001483:Base Sequence; D017931:DNA Primers / Q000737:chemistry; D004269:DNA, Bacterial / Q000032:analysis* / Q000737:chemistry; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D004927:Escherichia coli Infections / Q000382:microbiology; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D007525:Isoelectric Focusing; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D001618:beta-Lactamases / Q000235:genetics*
D017931:DNA Primers; D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S; D001618:beta-Lactamases