动物医学进展2024，Vol.45Issue(9)：45-48,4.
猪流行性腹泻病毒微芯片荧光定量RT-qPCR检测方法的建立
Establishment of Microchip RT-qPCR Method for Detection of Porcine Epidemic Diarrhea Virus
摘要
Abstract
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.关键词
猪流行性腹泻病毒/微芯片/实时荧光定量逆转录-聚合酶链反应/囊膜蛋白Key words
Porcine epidemic diarrhea virus/microchip/RT-qPCR/envelope protein分类
农业科技引用本文复制引用
曹洪志,王新杰,高姗姗,孙晓明,邓飞,石艳萍,陈弟诗,陈昌英,李平顺..猪流行性腹泻病毒微芯片荧光定量RT-qPCR检测方法的建立[J].动物医学进展,2024,45(9):45-48,4.基金项目
四川省科技计划重点研发项目(2023YFN0021) （2023YFN0021）
宜宾职业技术学院科技创新团队项目(ybzy21cxtd-08) （ybzy21cxtd-08）
宜宾职业技术学院自然科学研究项目(22ZRZD-09) （22ZRZD-09）
