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Loop-mediated isothermal amplification of vanA gene enables a rapid and naked-eye detection of vancomycin-resistant enterococci infection

Authors
 Hye Jin Kim  ;  Yu Jin Kim  ;  Dong Eun Yong  ;  Kyungwon Lee  ;  Jeon Han Park  ;  Jae Myun Lee  ;  Sang Sun Yoon 
Citation
 Journal of Microbiological Methods, Vol.104C : 61-66, 2014 
Journal Title
 Journal of Microbiological Methods 
ISSN
 0167-7012 
Issue Date
2014
Abstract
Vancomycin-resistant enterococci (VRE) are one of the leading causes of nosocomial infection at intensive care unit (ICU). A rapid and sensitive detection of VRE infection is in high demand for timely and suitable antibiotic treatment. Here, we optimized a distinct DNA-based diagnostic technique, loop-mediated isothermal amplification (LAMP) for a rapid detection of the presence of vanA gene, a critical component of the gene cluster required for vancomycin resistance. Amplification efficiency was optimal at 62 °C and with 2 mM MgSO4. The detection limit of the DNA template was 80 pg and LAMP amplicons were detected within 40 min; thereby suggesting a potential applicability of LAMP as a sensitive and urgent diagnostic method. Furthermore, positive LAMP reaction was directly detected with the naked-eye by monitoring the formation of a white precipitate or the color change induced by hydroxy naphthol blue (HNB) dye. Finally, 56 clinical isolates were successfully tested for the presence of vanA gene by LAMP, which was determined to be more sensitive than PCR. Together, our results clearly demonstrate the usefulness of LAMP for the diagnosis of VRE infection.
Full Text
http://www.sciencedirect.com/science/article/pii/S0167701214001511
DOI
10.1016/j.mimet.2014.05.021
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Laboratory Medicine (진단검사의학교실) > 1. Journal Papers
1. College of Medicine (의과대학) > Dept. of Microbiology (미생물학교실) > 1. Journal Papers
Yonsei Authors
박전한(Park, Jeon Han) ORCID logo https://orcid.org/0000-0001-9604-3205
용동은(Yong, Dong Eun) ORCID logo https://orcid.org/0000-0002-1225-8477
윤상선(Yoon, Sang Sun) ORCID logo https://orcid.org/0000-0003-2979-365X
이경원(Lee, Kyungwon) ORCID logo https://orcid.org/0000-0003-3788-2134
이재면(Lee, Jae Myun) ORCID logo https://orcid.org/0000-0002-5273-3113
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URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/99043
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