副溶血性弧菌恒温荧光法检测及快速检测试剂盒的初步应用

Real-time isothermal detection of Vibrio parahaemolyticus and its preliminary application in rapid detection kits

【目的】采用恒温荧光PCR方法及快速检测试剂盒对副溶血弧菌的扩增结果进行实时检测,并对其检测限、灵敏度及特异度等进行测试。【方法】通过三个实验阶段(实验室自配阳性样品测试阶段、腹泻病人模拟生物样本测试阶段、市场食品样测试阶段),验证仪器及试剂盒的稳定性、对腹泻样本和市场食品样本检测的可靠性,及其适用范围。【结果】实验室自配样品检测结果较好,检测限达到7~100 cfu/m L;腹泻样和市场食品样的检测假阳性率较高,但灵敏度极好,三个阶段合计灵敏度为99.6%,特异度为55.9%,真阳性率为85.1%,真阴性率为98.3%。【结论】该恒温荧光PCR方法及快速检测试剂盒灵敏度高,稳定性好,适用范围广,在控制实验操作过程及实验条件,提高特异度后,值得进一步推广,对我国食源性副溶血性弧菌的检测具有重要意义。

[Objective] To verify the limit of detection as well as sensitivity and specificity of real-time isothermal detection and rapid detection kit of Vibrio parahaemolyticus（ VP）. [Methods]By detecting three groups of samples-laboratory prepared samples,commercial food samples and diarrhea patients samples,the stability and reliability as well as the scope of application of the method and kit were verified. [Results] Laboratory configuration sample detection showed a stable and reliable result,in which the detection limit reached 7 to 100 cfu/m L. Although the FPR of food sample and diarrhea patients sample were relatively high,the sensitivity was great. The total sensitivity,specificity,true positive rate,true negative rate were respectively 99. 6%,55. 9%,85. 1%,98. 3%. [Conclusion]The real-time isothermal detection and rapid detection kit have a great sensitivity for VP detection. Also the results are stable and essentially in agreement with the culture method of GB 4789. 7—2013. The method is worth being promoted in application,under the premise that experimental operating steps and experimental conditions are wellcontrolled in order to improve the specificity.