Detection of mutations conferring extended-spectrum activity on SHV beta-lactamases using polymerase chain reaction single strand conformational polymorphism (PCR-SSCP)

J Antimicrob Chemother. 1996 Apr;37(4):797-802. doi: 10.1093/jac/37.4.797.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • DNA Mutational Analysis
  • Drug Resistance, Microbial
  • Humans
  • Isoelectric Focusing / methods
  • Molecular Sequence Data
  • Point Mutation / genetics*
  • Polymerase Chain Reaction / methods*
  • Polymorphism, Single-Stranded Conformational
  • beta-Lactamases / genetics*

Substances

  • beta-lactamase SHV-5
  • beta-Lactamases