Rapid detection and molecular survey of bla VIM, bla IMP and bla NDM genes among clinical isolates of Acinetobacter baumannii using new multiplex real-time PCR and melting curve analysis
BMC Microbiology(2019)
摘要
Background Acinetobacter baumannii is a cosmopolitan bacterium that is frequently reported from hospitalized patients, especially those patients who admitted in the intensive care unit. Recently, multiplex real-time PCR has been introduced for rapid detection of the resistance genes in clinical isolates of bacteria. The current study aimed to develop and evaluate multiplex real-time PCR to detect common resistance genes among clinical isolates of A. baumannii . Results Multiplex real-time PCR based on melting curve analysis showed different T m corresponding to the amplified fragment consisted of 83.5 °C, 93.3 °C and 89.3 °C for bla IMP, bla VIM and bla NDM, respectively. Results of multiplex real-time PCR showed that the prevalence of bla IMP, bla VIM and bla NDM among the clinical isolates of A. baumannii were 5/128(3.9%), 9/128(7.03%) and 0/128(0%), respectively. Multiplex real-time PCR was able to simultaneously identify the resistance genes, while showed 100% concordance with the results of conventional PCR. Conclusions The current study showed that bla VIM, was the most prevalent MBL gene among the clinical isolates of A. baumannii while no amplification of bla NDM was seen. Multiplex real-time PCR can be sensitive and reliable technique for rapid detection of resistance genes in clinical isolates.
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关键词
Acinetobacter baumannii,Melting curve analysis,Multiplex real-time PCR,Single tube reaction
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