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http://dx.doi.org/10.15616/BSL.2020.26.3.149

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

Baek, Yun Hee (Department of Microbiology, Chungbuk National University College of Medicine)
Hong, Seung Bok (Department of Clinical Laboratory Science, Chungbuk Health & Science University)
Shin, Kyeong Seob (Department of Laboratory Medicine, Chungbuk National University Hospital)
Publication Information
Biomedical Science Letters / v.26, no.3, 2020 , pp. 149-156 More about this Journal
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 (bioMérieux 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.
Keywords
Vancomycin-resistant Enterococci (VRE); Loop-mediated isothermal amplification; vanA; vanB; 16S rRNA;
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