Construction by polymerase chain reaction and use of intragenic DNA probes for three main types of transferable beta-lactamases (TEM, SHV, CARB) [corrected].
Intragenic DNA probes were synthesized by polymerase chain reaction using fragments of the genes of three major types of beta-lactamases (TEM, SHV, CARB) as templates. The TEM probe hybridized with the genes encoding TEM-1, TEM-2 and six extended-spectrum related enzymes (TEM-3 to TEM-7, TEM-2O) in colony hybridizations and Southern-blot analysis. The SHV probe hybridized with the genes for SHV-1, OHIO-1 and four derived extended-spectrum beta-lactamases (SHV-2, SHV-3, SHV-4 and SHV-5). The CARB probe hybridized with the genes for PSE-1 (CARB-2), PSE-4 (CARB-1), CARB-3 and CARB-4. None of the probes hybridized with genes for any of eight oxacillin-hydrolysing enzymes, PSE-2, OXA-1 to OXA-7, ROB-1 and chromosomal beta-lactamases of various Enterobacteriaceae (except Klebsiella pneumoniae) and Pseudomonas aeruginosa. Investigations of Escherichia coli clinical isolates using these probes indicate the presence of a novel type of extended-spectrum, transferable beta-lactamase.
Service de Bactériologie-Virologie, Hôpital Saint-Louis, Paris, France.
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D001483:Base Sequence; D015342:DNA Probes; D004352:Drug Resistance, Microbial / Q000235:genetics; D005798:Genes, Bacterial; D006090:Gram-Negative Bacteria / Q000235:genetics; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D011089:Polydeoxyribonucleotides / Q000235:genetics; D016133:Polymerase Chain Reaction; D011815:R Factors; D001618:beta-Lactamases / Q000145:classification / Q000235:genetics*
D015342:DNA Probes; D011089:Polydeoxyribonucleotides; D001618:beta-Lactamases