Rapid and sensitive detection of the vanA resistance gene from clinical Enterococcus faecium and Enterococcus faecalis isolates by loop-mediated isothermal amplification

Qian-Qian Huang; Bin-Bin Liu; Hui-Fen Zhu; Jian-Jia Ma; Madeleine Tsoi; Bing-Qing Yao; Li-Chen Yao; Qiang Wu; Xiao-Qin Mu; Shu-Lin Liu

doi:10.1016/j.jgar.2018.10.012

Rapid and sensitive detection of the vanA resistance gene from clinical Enterococcus faecium and Enterococcus faecalis isolates by loop-mediated isothermal amplification

J Glob Antimicrob Resist. 2019 Mar:16:262-265. doi: 10.1016/j.jgar.2018.10.012. Epub 2018 Oct 16.

Authors

Affiliations

¹ Systemomics Center, College of Pharmacy, and Genomics Research Center, Harbin Medical University, Harbin, China; HMU-UCCSM Center for Infection and Genomics, Harbin Medical University, Harbin, China; Translational Medicine Research and Cooperation Center of Northern China, Heilongjiang Academy of Medical Sciences, Heilongjiang, China.
² Systemomics Center, College of Pharmacy, and Genomics Research Center, Harbin Medical University, Harbin, China; HMU-UCCSM Center for Infection and Genomics, Harbin Medical University, Harbin, China; Translational Medicine Research and Cooperation Center of Northern China, Heilongjiang Academy of Medical Sciences, Heilongjiang, China. Electronic address: muxiaoqin@ems.hrbmu.edu.cn.
³ Systemomics Center, College of Pharmacy, and Genomics Research Center, Harbin Medical University, Harbin, China; HMU-UCCSM Center for Infection and Genomics, Harbin Medical University, Harbin, China; Translational Medicine Research and Cooperation Center of Northern China, Heilongjiang Academy of Medical Sciences, Heilongjiang, China; Department of Microbiology, Immunology and Infectious Diseases, University of Calgary, Calgary, Canada. Electronic address: slliu@hrbmu.edu.cn.

PMID: 30339895
DOI: 10.1016/j.jgar.2018.10.012

Abstract

Objectives: Vancomycin resistance in Enterococcus spp., mediated mainly by the vanA resistance gene, has become a major health concern as it has spread worldwide. Therefore, a rapid method is urgently required to detect the vanA gene for timely and appropriate antimicrobial control of resistant Enterococcus infections.

Methods: The loop-mediated isothermal amplification (LAMP) assay was optimised for vanA detection in Enterococcus spp. isolates.

Results: The LAMP primer set designed in this study could reliably recognise seven distinct regions of the vanA gene and amplify the gene within 25min at an isothermal temperature of 65°C with high specificity. The sensitivity of the optimised assay was high, with a detection limit for vanA as low as 100pg/μL, which is 100-fold more sensitive than the PCR assay. A special advantage of this optimised LAMP method is that the vanA gene could be detected directly from clinical specimens.

Conclusion: This optimised LAMP assay has great application potential for efficient detection of vanA in clinical diagnosis and epidemiological studies.

Keywords: Antimicrobial resistance; LAMP; Vancomycin-resistant Enterococcus; vanA.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adolescent
Adult
Anti-Bacterial Agents / pharmacology*
Bacterial Proteins / genetics*
Carbon-Oxygen Ligases / genetics*
Enterococcus faecalis / drug effects*
Enterococcus faecalis / genetics*
Enterococcus faecalis / isolation & purification
Enterococcus faecium / drug effects*
Enterococcus faecium / genetics*
Enterococcus faecium / isolation & purification
Female
Humans
Male
Microbial Sensitivity Tests
Middle Aged
Nucleic Acid Amplification Techniques*
Sensitivity and Specificity
Temperature
Vancomycin Resistance / genetics
Young Adult

Substances

Anti-Bacterial Agents
Bacterial Proteins
VanA ligase, Bacteria
Carbon-Oxygen Ligases