Construction of standard plasmids for detecting Salmonella in meat products with real-time fluorescence quantitative polymerase chain reaction
- VernacularTitle:肉制品中沙门菌实时荧光定量聚合酶链反应标准质粒的构建
- Author:
Ying YU
1
;
Qi-Fang GU
;
Min CHEN
;
Hong-Zhi ZHANG
Author Information
1. 上海市疾病预防控制中心
- Keywords:
plasmid standard;
fluorescence quantitative polymerase chain reaction;
Salmonella;
meat product
- From:
Shanghai Journal of Preventive Medicine
2017;29(4):273-276,280
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct standard plasmids for detecting Salmonella in meat products with real-time fluorescence quantitative polymerase chain reaction(PCR).Methods Primers directed at Salmonella invA gene were designed.Specific fragments were amplified and cloned into plasmid vectors to construct recombinant plasmids.The constructed recombinant plasmids were used as standards for implementing the real-time fluorescence quantitative PCR after optimization, establishing standard curves and examining the sensitivity and stability of these standards.Results The plasmid standard containing the invA gene in Salmonella was successfully constructed.The cycle threshold value(Ct value) of the standard curve established by means of the plasmid standard and the number of template copies exhibited good linear relationship(r2=0.9979).The minimum that could be detected by means of this method was 10 copies/reaction.The standard plasmid was proved to have good stability.This standard plasmid was used to detect Salmonella in meat products.After two hours enrichment, sample detection could be completed within seven hours.Conclusion The constructed plasmid standard can be used for detecting Salmonella in meat products by means of the fluorescence quantitative PCR, which can provide reliable reference bases for examining quality of relevant experiments.
