학술논문
H7N9亚型 AIV 双重实时荧光定量 RT-PCR 检测方法的建立 / Development of a Duplex Real-time PCR for Detection of H7N9 Avian Influenza Virus
Document Type
Academic Journal
Author
罗思思; 谢芝勋; 刘加波; 陈敏玫; 杨进业; 邓显文; 谢志勤; 庞耀珊; 谢丽基; 范晴; LUO Si-si; XIE Zhi-xun; LIU Jia-bo; CHEN Min-mei; YANG Jin-ye; DENG Xian-wen; XIE Zhi-qin; PANG Yao-shan; XIE Li-ji; FAN Qing
Source
动物医学进展 / Progress in Veterinary Medicine. (12):1-5
Subject
Language
Chinese
ISSN
1007-5038
Abstract
根据 H7N9亚型禽流感病毒（AIV）HA 基因和 NA 基因的保守序列，分别设计特异性引物和不同荧光基团标记的 TaqMan 探针。通过优化反应条件，建立了 H7N9亚型 AIV 的双重实时荧光定量 RT-PCR 检测方法。结果显示，该法检测 H7N9亚型 AIV 的下限为102 copies/μL，批内重复和批间重复变异系数均小于3％。本研究建立的 H7N9亚型 AIV 双重实时荧光定量 RT-PCR 方法，具有快速、特异和敏感的优点，可为 H7N9亚型 AIV 的有效防控提供技术支撑。
The specific primers and TaqMan probes were designed according to the conserved sequences of the hemagglutinin (HA)genes of H7 subtype AIV and the neuramidinase(NA)genes of N9 subtype AIV, respectively.The reaction conditions were optimized to develop a duplex real-time RT-PCR assay for rapid detection of H7N9 AIV.It was shown that the specificity of this assay was high,the amplification curves were displayed for the detection of H7N9 AIV,H7 subtype AIV and N9 subtype AIV,and no positive re-sults were observed when nucleic acid from other subtypes AIV and avian pathogens.The detection limit of this assay was 100 copies of H7N9 AIV,and the coefficients of variation were both less than 3% for the intra-assay and inter-assay.This duplex assay is a rapid,specific and sensitive method for the detection of H7N9 AIV,and provides a technical support to prevent and control H7N9 AIV.
The specific primers and TaqMan probes were designed according to the conserved sequences of the hemagglutinin (HA)genes of H7 subtype AIV and the neuramidinase(NA)genes of N9 subtype AIV, respectively.The reaction conditions were optimized to develop a duplex real-time RT-PCR assay for rapid detection of H7N9 AIV.It was shown that the specificity of this assay was high,the amplification curves were displayed for the detection of H7N9 AIV,H7 subtype AIV and N9 subtype AIV,and no positive re-sults were observed when nucleic acid from other subtypes AIV and avian pathogens.The detection limit of this assay was 100 copies of H7N9 AIV,and the coefficients of variation were both less than 3% for the intra-assay and inter-assay.This duplex assay is a rapid,specific and sensitive method for the detection of H7N9 AIV,and provides a technical support to prevent and control H7N9 AIV.