Development of multiplex loop-mediated isothermal amplification (mLAMP) for detection of Salmonella,Vibrio parahaemolyticus and Listeria monocytogenes
10.7644/j.issn.1674-9960.2016.09.018
- VernacularTitle:多重环介导等温扩增快速检测沙门菌、副溶血弧菌和单核细胞增生性李斯特菌方法的建立
- Author:
Ningwei LIU
;
Dayang ZOU
;
Derong DONG
;
Zhan YANG
;
Simei HUANG
;
Xiaoming HE
;
Da AO
;
Wei LIU
;
Liuyu HUANG
- Publication Type:Journal Article
- Keywords:
multiplex loop-mediated isothermal amplification;
Salmonella;
Vibrio parahaemolyticus;
Listeria monocyto-genes
- From:
Military Medical Sciences
2016;40(9):767-772
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a multiplex loop-mediated isothermal amplification(mLAMP)method for simultaneous detection of Salmonella,Vibrio parahaemolyticus (VPH)and Listeria monocytogenes (LM).Methods Three sets of mLAMP primers were designed to specifically target bcfD of Salmonella and tlh of VPH and iap of LM.The respective single LAMP assay of the three kinds of bacteria was developed,and the ratio of primer concentration was optimized to develop a multiplex LAMP system.The specificity and sensitivity of multiplex LAMP were observed.Results Turbidity monitoring results in real time suggests that the mLAMP was highly specific and amplification could be obtained within 45 min under isothermal conditions.The sensitivity of this mLAMP was found to be 300 fg/μl genomic DNAs for Salmonella and 4.2 pg/μl for VPH and 4.5 pg/μl for LM,which was consistent with conventional PCR.Conclusion The mLAMP described can potentially facilitate simultaneous detection of three kinds of bacteria in a large number of food samples, which could be used as a primary screening method and as a supplement to classical detection methods.
