Rapid detection and molecular survey of blaVIM, blaIMP and blaNDM genes among clinical isolates of Acinetobacter baumannii using new multiplex real-time PCR and melting curve analysis

被引:17
作者
Goudarzi, Hossein [1 ]
Mirsamadi, Elnaz Sadat [1 ,2 ]
Ghalavand, Zohreh [1 ]
Vala, Mojdeh Hakemi [1 ]
Mirjalali, Hamed [3 ]
Hashemi, Ali [1 ]
机构
[1] Shahid Beheshti Univ Med Sci, Dept Microbiol, Fac Med, Tehran, Iran
[2] Islamic Azad Univ, Dept Microbiol, Fac Med, Tehran Med Sci, Tehran, Iran
[3] Shahid Beheshti Univ Med Sci, Res Inst Gastroenterol & Liver Dis, Foodborne & Waterborne Dis Res Ctr, Tehran, Iran
关键词
Acinetobacter baumannii; Melting curve analysis; Multiplex real-time PCR; Single tube reaction; GRAM-NEGATIVE PATHOGENS; MULTIDRUG-RESISTANT; ANTIBIOTIC-RESISTANCE; CARBAPENEMASE GENES; EPIDEMIOLOGY; IDENTIFICATION; MECHANISMS;
D O I
10.1186/s12866-019-1510-y
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
BackgroundAcinetobacter 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.ResultsMultiplex real-time PCR based on melting curve analysis showed different T-m corresponding to the amplified fragment consisted of 83.5 degrees C, 93.3 degrees C and 89.3 degrees C for blaIMP, blaVIM and blaNDM, respectively. Results of multiplex real-time PCR showed that the prevalence of blaIMP, blaVIM and blaNDM 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.ConclusionsThe current study showed that blaVIM, was the most prevalent MBL gene among the clinical isolates of A. baumannii while no amplification of blaNDM 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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