猪轮状病毒TaqMan荧光定量检测方法的建立及初步应用
Establishment and preliminary application of TaqMan fluorescence quantitative detection method for porcine rotavirus
林正丹 1涂军 2詹存林 1孙秀秀 1冯贺龙 1刘茜 1胡薛英 1谷长勤 1张万坡 1陶攀 1陈品 1余腾 2钱平 1程国富1
作者信息
- 1. 华中农业大学动物医学学院,湖北武汉 430070
- 2. 广西扬翔股份有限公司,广西 贵港 537000
- 折叠
摘要
为了对早期猪轮状病毒(porcine rotavirus,PoRV)感染情况进行监测,根据GenBank中已有的A型PoRV VP6序列,设计2对引物和荧光定量探针,建立了 TaqMan荧光定量RT-PCR方法,并对该方法特异性、灵敏性和重复性进行评价.结果显示所建立的TaqMan荧光定量RT-PCR方法特异性强、重复性好、灵敏度高,可检测到101拷贝/μL.经对2 366份腹泻仔猪的粪便样品进行检测,所建立的方法检出率(5.92％,140/2 366)高于常规RT-PCR(4.95％,117/2 366),两者的符合率为98.58％.结果表明,本研究建立的PoRV TaqMan荧光定量RT-PCR方法可用于猪轮状病毒病的临床检测.
Abstract
In order to monitor the early porcine rotavirus infection,according to the existing porcine rotavirus A VP6 sequence in GenBank,two pairs of primers and fluorescence probes were de-signed,and the TaqMan real-time RT-PCR method was established,and the specificity,sensitivity and repeatability of the method were evaluated.The results showed that the established TaqMan real-time RT-PCR method had strong specificity,good repeatability and high sensitivity,and could detect 101 copies/μL.After testing the fecal samples of 2 366 diarrheal piglets,the detection rate of the established method(5.92％,140/2 366)was higher than that of conventional RT-PCR(4.95％,117/2 366),and the compliance rate of the two was 98.58％.The results showed that the TaqMan fluorescence RT-PCR method established in this study can be used for the clinical detec-tion of porcine rotavirus disease.
关键词
猪轮状病毒/TaqMan探针/荧光定量RT-PCR方法Key words
porcine rotavirus/TaqMan probe/quantitative RT-PCR method引用本文复制引用
基金项目
中央高校基本科研业务费专项(140422008)
扬翔股份有限公司重大创新变革基金(JYKJ-R&D2022-01)
出版年
2024
NETL
NSTL
万方数据