Quantitative multiplex real-time PCR for detecting class 1, 2 and 3 integrons. - Inserm - Institut national de la santé et de la recherche médicale Access content directly
Journal Articles Journal of Antimicrobial Chemotherapy Year : 2010

Quantitative multiplex real-time PCR for detecting class 1, 2 and 3 integrons.

Abstract

OBJECTIVES: 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 probe-based real-time PCR method with three different primer-probe pairs for simultaneous detection of the three main classes of integron. METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.

Dates and versions

inserm-00533199 , version 1 (05-11-2010)

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Olivier Barraud, Marie-Claire Baclet, François Denis, Marie-Cécile Ploy. Quantitative multiplex real-time PCR for detecting class 1, 2 and 3 integrons.. Journal of Antimicrobial Chemotherapy, 2010, 65 (8), pp.1642-1645. ⟨10.1093/jac/dkq167⟩. ⟨inserm-00533199⟩
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