基于双重数字PCR检测副溶血弧菌和鼠伤寒沙门氏菌

Detection of Vibrio Parahaemolyticus and Salmonella Typhimurium by Duplex Droplet Digital PCR

本研究利用非特异性核酸染料Eva Green,建立检测副溶血弧菌Tlh基因和鼠伤寒沙门氏菌InvA基因的双重数字PCR方法,并分析该检测方法的特异性、灵敏度及重复性。结果显示,所建立的方法对副溶血弧菌和鼠伤寒沙门氏菌的检测具有较高的特异性和灵敏度,检测DNA最低浓度为10 fg·μL^(-1),tlh最低检测限为16.8 copies/20μL,invA最低检测限为8.6 copies/20μL,该方法特异性强、灵敏度高且重复性好,可用于食品中副溶血弧菌和鼠伤寒沙门氏菌的检测。

In this study,the non-specific nucleic acid dye Eva Green was used to establish a duplex digital PCR method to detect the Tlh gene of Vibrio parahaemolyticus and the Inv A gene of Salmonella typhimurium,and the specificity,sensitivity and repeatability of the detection method were analyzed.The results showed that the developed method could specifically detect Vibrio parahaemolyticus and Salmonella typhimurium with a detectable DNA template concentration of 10 fg·μL^(-1).The limit of detection for tlh was 16.8 copies/20μL,and the limit of detection for invA is 8.6 copies/20μL.The method has strong specificity,high sensitivity and good reproducibility,and can be used for the detection of Vibrio parahaemolyticus and Salmonella typhimurium.