Detection of TEM beta-lactamase genes by non-isotopic spot hybridisation.
Eur J Clin Microbiol
; 6(4): 406-9, 1987 Aug.
Article
em En
| MEDLINE
| ID: mdl-3311735
ABSTRACT
A 998 bp fragment of plasmid pBR322, comprising part of the TEM-1 beta-lactamase gene, was labelled with biotin-11-dUTP for use as a DNA probe in a rapid non-isotopic spot hybridisation test. Diluted broth cultures of bacteria producing different beta-lactamases were filtered onto nitrocellulose and lysed in situ. Following pre-hybridisation treatment with proteinase K, hybridisation with the labelled probe was demonstrated using a commercially available streptavidine/polyalkaline phosphatase-based detection system. The probe was highly specific, reacting only with strains producing either the TEM-1 or structurally similar TEM-2 enzyme. An inoculum of 3-4 X 10(6) cells gave optimum positive discrimination. When 90 recent ampicillin-resistant strains of Escherichia coli isolated from patients with urinary tract infections were screened using the system, 72% gave a positive hybridisation signal.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Tipo de estudo:
Diagnostic_studies
Limite:
Humans
Idioma:
En
Ano de publicação:
1987
Tipo de documento:
Article