A rapid loop-mediated isothermal amplification (LAMP) method for detection of the macrolide-streptogramin type B resistance gene msrA in Staphylococcus aureus.
Macrolide-streptogramin type B resistance (the MSB phenotype) is a multidrug resistance phenotype in Staphylococcus aureus conferred by the resistance gene msrA. However, bacteria having the MSB phenotype are susceptible to lincosamides and 16-membered ring macrolides, which makes profiling resistance genes necessary and urgent for timely and appropriate use of antimicrobials. In this study, the loop-mediated isothermal amplification (LAMP) assay was optimized for prompt detection of the msrA gene. msrA gene sequences were obtained from the National Center for Biotechnology Information (NCBI) database and primers were designed using the LAMP primer designing software PrimerExplorer v.4, which together recognize seven distinct regions of the msrA gene. The specific LAMP primer set designed in this study could amplify the msrA gene within 25min at an isothermal temperature of 62°C. More importantly, the msrA gene could be detected at a sensitivity as low as 100pg. Furthermore, this optimized LAMP assay provided swift detection of the msrA gene even directly from human specimens. In conclusion, this assay may have great clinical application potential for detection of the msrA gene.
Systemomics Center, College of Pharmacy, and Genomics Research Center (State-Province Key Laboratories of Biomedicine-Pharmaceutics of China), Harbin Medical University, Harbin, China.
Xiao-Qin
XQ
Mu
Systemomics Center, College of Pharmacy, and Genomics Research Center (State-Province Key Laboratories of Biomedicine-Pharmaceutics of China), Harbin Medical University, Harbin, China.
Bin-Bin
BB
Liu
Systemomics Center, College of Pharmacy, and Genomics Research Center (State-Province Key Laboratories of Biomedicine-Pharmaceutics of China), Harbin Medical University, Harbin, China; HMU-UCFM Centre for Infection and Genomics, Harbin Medical University, Harbin, China.
Ephraim
E
Hui
Systemomics Center, College of Pharmacy, and Genomics Research Center (State-Province Key Laboratories of Biomedicine-Pharmaceutics of China), Harbin Medical University, Harbin, China; HMU-UCFM Centre for Infection and Genomics, Harbin Medical University, Harbin, China.
William
W
Huang
Systemomics Center, College of Pharmacy, and Genomics Research Center (State-Province Key Laboratories of Biomedicine-Pharmaceutics of China), Harbin Medical University, Harbin, China.
Li-Chen
LC
Yao
Department of Medicine Laboratory, The Second Affiliated Hospital of Harbin Medical University, Harbin, China.
Li-Bo
LB
Duo
Department of Medicine Laboratory, The Second Affiliated Hospital of Harbin Medical University, Harbin, China.
Wen-Ying
WY
Sun
Department of Laboratory Diagnosis, The First Affiliated Hospital of Harbin Medical University, Harbin, China.
Gui-Qiu
GQ
Li
Department of Infectious Diseases, The Fourth Affiliated Hospital of Harbin Medical University, Harbin, China.
Fu-Xiang
FX
Wang
Systemomics Center, College of Pharmacy, and Genomics Research Center (State-Province Key Laboratories of Biomedicine-Pharmaceutics of China), Harbin Medical University, Harbin, China; HMU-UCFM Centre for Infection and Genomics, Harbin Medical University, Harbin, China; Department of Microbiology, Immunology and Infectious Diseases, University of Calgary, Calgary, Canada. Electronic address: slliu@hrbmu.edu.cn.
Shu-Lin
SL
Liu
D017931:DNA Primers; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D005784:Gene Amplification; D005798:Genes, Bacterial; D006801:Humans; D055231:Lincosamides; D018942:Macrolides; D021141:Nucleic Acid Amplification Techniques; D012680:Sensitivity and Specificity; D013203:Staphylococcal Infections; D013211:Staphylococcus aureus / Q000235:genetics*; D025381:Streptogramin B
D017931:DNA Primers; D055231:Lincosamides; D018942:Macrolides; D025381:Streptogramin B
Antimicrobial resistance; Diagnostics; LAMP; Staphylococcus aureus; msrA gene