引物间隔臂修饰对液相芯片食源性致病菌检测的影响

Effect of Spacer Modified Primers on Detection of Food Borne Pathogens in Liquid Chip

建立液相芯片同时检测鼠伤寒沙门氏菌(S.typhimurium)、志贺氏菌(Shigella)方法,并对引物进行间隔臂(Spacer C3)修饰,验证C3修饰对食源性致病菌检测的影响。对引物C3修饰组和未修饰组两种食源性致病菌进行条件优化和对比。试验表明所建立的食源性致病菌检测体系方法学评价良好,多批次结果显示稳定性较好,反应体系退火温度和杂交温度分别确定为56℃和38℃。结果表明,引物使用C3修饰对检测结果的阳性荧光值强度中值(median fluorescence intensity,MFI)有良好信号放大作用,为食源性致病菌液相芯片检测提供了参考。

A liquid phase chip was established to simultaneously detect S.typhimurium and Shigella,and the primers were modified by Spacer C3 to verify the effect of C3 modification on the detection of food-borne pathogens.Two kinds of food-borne pathogens were modified and compared between the C3 modified group and the unmodified group.The test showed that the established foodborne pathogen detection system had a good methodological evaluation.The results of multiple batches showed good stability.The annealing temperature and hybridization temperature of the reaction system were determined to be 56℃and 38℃,respectively.The results showed that the primers modified by C3 had a good signal amplification effect on the median fluorescence intensity of the detection results,which provided a reference for the detection of foodborne pathogenic bacteria in the liquid phase chip.