蜜蜂慢性麻痹病毒荧光定量RT-PCR检测方法的建立
Development of a Real-Time RT-PCR Method for Detection of Chronic Bee Paralysis Virus
张体银 1王武军 1林素洁 1张志灯 1李宋钰 1于师宇1
作者信息
- 1. 福州海关技术中心福建省检验检疫技术研究重点实验室,福州 350003
- 折叠
摘要
为建立慢性蜜蜂麻痹病毒(CBPV)快速诊断方法,本研究根据CBPV RNA依赖RNA聚合酶(RdRp)基因保守区设计特异性引物和TaqMan探针,建立了荧光定量RT-PCR检测方法.结果显示,以构建的重组质粒为标准品建立的TaqMan荧光定量PCR方法,标准曲线具有良好的线性关系,线性相关系数达0.998;该方法最低检出限为10拷贝/μL,与蜜蜂急性麻痹病毒等常见蜜蜂病毒无交叉反应,具有良好的灵敏性和特异性;组内和组间变异系数分别低于0.5％和2％,具有较好的稳定性.本研究建立的CBPV荧光定量RT-PCR检测方法,可用于实验室检测、流行病学调查和疫情监测.
Abstract
To establish a rapid diagnostic method for chronic bee paralysis virus(CBPV),a fluorogenic TaqMan real-time PCR assay was developed by designing specific primers and TaqMan probes according to the RNA dependent RNA polymerase(RdRp)gene sequence of CBPV in this study.The results showed that the real-time RT-PCR method had a good linear relation,and linear correlation coefficient R2 was 0.998.The sensitivity limit was 10 copies/μL and there was no cross-reaction with other Honey bee virus,which indicated that the method had good sensitivity and specificity.The coefficients of variation(CV)for intra-assay and inter-assay repeatability were less than 0.5％and 2％,respectively,showing that the method had good stability.The real-time RT-PCR method for CBPV developed in this study might be used for laboratory testing,epidemiological investigation and epidemic monitoring.
关键词
蜜蜂慢性麻痹病毒/荧光定量RT-PCR/RNA依赖RNA聚合酶Key words
Chronic bee paralysis virus/real-time RT-PCR/RNA dependent RNA polymerase(RdRp)引用本文复制引用
基金项目
海关总署科研项目(2021HK161)
福建省对外合作重点项目(202110029)
福州海关科研项目(FK2020-21)
出版年
2024
国家科技期刊平台
NSTL
万方数据