猪流行性腹泻病毒微芯片荧光定量RT-qPCR检测方法的建立OA北大核心CSTPCD

In order to establish a fluorescent quantitative reverse transcription-polymerase chain reaction(RT-qPCR)assay for porcine epidemic diarrhea virus(PEDV)with easy operation and rapidity,specific primers and probes were designed based on the published PEDV vesicular membrane protein E gene in the GenBank database combined with the microchip technology,and a microchip RT-qPCR assay was estab-lished.The method was positive for PEDV and negative for other disease-causing viruses with close homol-ogy,and the lowest detection limit was 1 copies/μL,while the detection limit of conventional RT-qPCR was 10 copies/μL.The intra-and inter-batch reproducibility experiments showed that the coefficient of variation of Ct value was within 2％,and the method was able to detect clinical-positive samples of PEDV.A novel microchip RT-qPCR assay of PEDV with good specificity,high sensitivity,short detection time and good reproducibility was established,which is suitable for clinical testing.