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

• Published in: Journal of antimicrobial chemotherapy 1996-04, Vol.37 (4), p.797-802
• Main Authors: M'Zali, Fatima-Hannachi, Gascoyne-Binzi, Deborah M., Heritage, John, Hawkey, Peter M.
• Format: Article
• Language: English
• Subjects: Bacteriological methods and techniques used in bacteriology; Bacteriology; Base Sequence; beta-Lactamases - genetics; Biological and medical sciences; DNA Mutational Analysis; Drug Resistance, Microbial; Fundamental and applied biological sciences. Psychology; Humans; Isoelectric Focusing - methods; Microbiology; Molecular Sequence Data; Point Mutation - genetics; Polymerase Chain Reaction - methods; Polymorphism, Single-Stranded Conformational
• ISSN: 0305-7453; 1460-2091
• DOI: 10.1093/jac/37.4.797

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 β-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 SHY β-lactamases.