期刊
JOURNAL OF ANTIMICROBIAL CHEMOTHERAPY
卷 65, 期 8, 页码 1642-1645出版社
OXFORD UNIV PRESS
DOI: 10.1093/jac/dkq167
关键词
qPCR; antibiotic resistance; genotypic detection; molecular diagnostic techniques
资金
- Ministere de l'Enseignement Superieur et de la Recherche
- Conseil Regional du Limousin
- Institut National de la Sante et de la Recherche Medicale (INSERM)
Integrons are bacterial genetic elements that can capture and express genes contained in mobile cassettes. Integrons have been described worldwide in Gram-negative bacteria and are a marker of antibiotic resistance. We developed a specific and sensitive Taqman((R)) probe-based real-time PCR method with three different primer-probe pairs for simultaneous detection of the three main classes of integron. Sensitivity was assessed by testing mixtures of the three targets (intI integrase genes of each integron class) ranging from 10 to 10(8) copies. Specificity was determined with a panel of integron-containing and integron-free control strains. The method was then applied to clinical samples. The PCR method was specific and had a sensitivity of 10(2) copies for all three genes, regardless of their respective quantities. The method was quantitative from 10(3) to 10(7) copies, and was able to detect integrons directly in biological samples. We have developed a rapid, quantitative, specific and sensitive method that could prove useful for initial screening of Gram-negative isolates, or clinical samples, for likely multidrug resistance.
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