Research and development of genetic diagnostic method of Staphyloccocus aureus based on loop-mediated isothermal amplification
10.3760/cma.j.issn.0254-5101.2010.04.022
- VernacularTitle:使用环介导等温扩增技术快速检测金黄葡萄球菌的初步研究
- Author:
Haihua YI
;
Guanghui HE
;
Chao FANG
;
Yangwei SONG
;
Bo XU
;
Huiyu SUN
;
Yunfei WANG
;
Wei WANG
;
Zheng XU
;
Jinwei ZHAO
- Publication Type:Journal Article
- Keywords:
Staphyloccocus aureus;
Loop-mediated isothermal amplification;
Rapid detection
- From:
Chinese Journal of Microbiology and Immunology
2010;30(4):382-386
- CountryChina
- Language:Chinese
-
Abstract:
Objective To develop a method of loop-mediated isothermal amplification(LAMP) to Staphyloccocus aureus rapidly, specifically, sensitively and simply suited for the primary health agency. Methods According to conserved nucleotide of Staphyloccocus aureus and principle of LAMP, we designed a set of LAMP primers and set up an LAMP reaction system. We evaluated the specificity, sensitivity and re-peatability of the method. In addition,we evaluated the linearity between initial template copies 1g value and reaction time (the time when the fluorescent value is 1×10~4). Results The optimal assay showed that it was no cross-reaction with other closely related members of pathogens, and was 10 times more sensitive than PCR. The coefficient of variance between tests was less than 5% ,and the kinetics curves showed a good line-arity between initial template copies lg value and reaction time(r~2=0. 9501). The detection activity could be finished within 1 h with the sensitivity of LAMP was 100% and the specificity was 94.4%, and the accuracy was 96.6%. Conclusion These findings demonstrated that the LAMP had the potential clinical application for detection and differentiation of Staphyloccocus aureus in the public health agency for its sensitive, specific and simple feature.
