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Simple and Rapid Detection of Vancomycin-Resistance Gene from Enterococci by Loop-Mediated Isothermal Amplification

Biomedical Science Letters 2020년 26권 3호 p.149 ~ 156
백윤희, 홍승복, 신경섭,
소속 상세정보
백윤희 ( Baek Yun-Hee ) - Chungbuk National University College of Medicine Department of Microbiology
홍승복 ( Hong Seung-Bok ) - Chungbuk Health and Science University Department of Clinical Laboratory Science
신경섭 ( Shin Kyeong-Seob ) - Chungbuk National University Hospital Department of Labaratory Medicine

Abstract


We developed a simple and rapid method for detecting vancomycin resistance genes, such as vanA and vanB, using loop-mediated isothermal amplification (LAMP). To identify not only vancomycin resistance genes, but also the genus Enterococcus, primers were designed for vanA, vanB, and 16S rRNA. Screening for vancomycin susceptibility in Enterococcus was performed using Etest (bioMerieux Inc). The results of the LAMP assay were compared to those of real-time RT-PCR. The optimal conditions for the LAMP assay were 65℃ for 60 min. The detection limits of the LAMP assay for vanA, and vanB were 2 × 102 copies/reaction. Compared to RT-PCR, the sensitivities and specificities of LAMP for 16S rRNA, vanA, and vanB were 100/100%, 100/100%, and 100/100%, respectively. The vanA genotype-vanB phenotype accounted for 57.5% (46/80) of the vancomycin-resistant Enterococci samples collected from 2016 to 2019. In conclusion, the LAMP assay developed in this study showed high sensitivity and specificity for vancomycin-resistant genes. Moreover, due to the simplicity and rapidity of the LAMP assay, its use can be very useful in clinical microbiology laboratories.

키워드

Vancomycin-resistant Enterococci (VRE); Loop-mediated isothermal amplification; vanA; vanB; 16S rRNA

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