Percoll分离法应用于肉制品中沙门菌核酸快速检测

Rapid detection of Salmonella nucleic acid in meat product by Percoll

目的用Percoll分离法提取核酸检测肉制品中沙门菌,提高肉制品中沙门菌核酸检测速度。方法用Percoll分离液从不同增菌时间的增菌液中提取沙门菌DNA进行PCR扩增,与煮沸裂解法提取核酸的PCR结果进行比较,并进行沙门菌定量计数。分析PCR扩增Ct值及扩增曲线形态。结果用Percoll分离法检测肉制品中沙门菌方法,可以在增菌9 h后得到扩增信号,增菌14 h可得到阳性检测结果,Percoll分离液中菌体的回收率仍有待提高。结论 Percoll分离液是一种良好的分离液,可有效去除肉制品中的基质,减少PCR扩增抑制因素,提高沙门菌核酸质量,减少肉制品中沙门菌引起食物中毒致病因子的检测时间。Percoll分离液中菌体的回收技术仍有待优化改进。

Objective To extract nucleic acid of Salmonella in meat product by the method of Percoll, so as to improve the de- tection speed of Salmonella nucleic acid in meat product. Methods Using separation liquid Percoll, to extract Salmonella DNA from different enrichment broth with different enrichment time for amplification by PCR, and compare with the PCR results with the boiling lysis nucleic acid for extraction, and Salmonella quantitative count was conducted. Analysis of PCR amplification of Ct value and the amplification curve shape were performed. Results Using Percoll to detect Salmonella in meat product, we can get amplification signals in 9 hours and get positive PCR results in 14 hours. The recovery rate of the bacteria by percoll need to be improved. Conclusion Percoll separation liquid is good. It can remove the food stroma and reduce the amplification inhibitory factor, and reduce the time for detection of Salmonella in meat product by promote the quality of nucleic acid. The re- covery rate of the bacteria by percoll still needs to be improved.