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

 Hye Jin Kim  ;  Yu Jin Kim  ;  Dong Eun Yong  ;  Kyungwon Lee  ;  Jeon Han Park  ;  Jae Myun Lee  ;  Sang Sun Yoon 
 Journal of Microbiological Methods, Vol.104C : 61-66, 2014 
Journal Title
 Journal of Microbiological Methods 
Issue Date
Bacterial Proteins/genetics* ; Carbon-Oxygen Ligases/genetics* ; Cross Infection/microbiology* ; Feces/microbiology ; Gram-Positive Bacterial Infections/microbiology* ; Humans ; Nucleic Acid Amplification Techniques/methods* ; Vancomycin-Resistant Enterococci/enzymology* ; Vancomycin-Resistant Enterococci/genetics ; Vancomycin-Resistant Enterococci/isolation & purification*
Diagnosis ; Loop-mediated isothermal amplification (LAMP) ; Vancomycin-resistant enterococci (VRE)
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.
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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
Park, Jeon Han(박전한) ORCID logo https://orcid.org/0000-0001-9604-3205
Yong, Dong Eun(용동은) ORCID logo https://orcid.org/0000-0002-1225-8477
Yoon, Sang Sun(윤상선) ORCID logo https://orcid.org/0000-0003-2979-365X
Lee, Kyungwon(이경원) ORCID logo https://orcid.org/0000-0003-3788-2134
Lee, Jae Myun(이재면) ORCID logo https://orcid.org/0000-0002-5273-3113
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