Detection of mutations conferring extended-spectrum activity on SHV beta-lactamases using polymerase chain reaction single strand conformational polymorphism (PCR-SSCP).
Single strand conformational polymorphism (SSCP) is a recently developed technique used to detect single base mutations in short PCR-generated amplimers. The method has been adapted and applied to differentiation of beta-lactamase genes. Each of the five standard SHV strains used produced a unique SSCP pattern, allowing the possibility of rapid identification of the SHV genes of other isolates. A clinical isolate that phenotypically produced SHV-5 yielded a pattern of major bands indistinguishable from that of the SHV-5 standard strain, illustrating the applicability of this technique. We therefore report a reliable and reproducible technique that can be applied to the characterisation of the SHV beta-lactamases.
M'Zali
F H
FH
Department of Microbiology, University of Leeds, UK.
Gascoyne-Binzi
D M
DM
Heritage
J
J
Hawkey
P M
PM
D001483:Base Sequence; D004252:DNA Mutational Analysis; D004352:Drug Resistance, Microbial; D006801:Humans; D007525:Isoelectric Focusing / Q000379:methods; D008969:Molecular Sequence Data; D017354:Point Mutation / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D018807:Polymorphism, Single-Stranded Conformational; D001618:beta-Lactamases / Q000235:genetics*
C117193:beta-lactamase SHV-5; D001618:beta-Lactamases