猪塞内卡病毒(SVA)和O型口蹄疫病毒双重荧光RT-PCR检测方法中阳性参考品的制备  

Preparation of Positive Reference in Duplex Fluorescent RT-PCR for detceting SVA and FMDV-O

作　　者：陈信全 张展 周伟光 张峥嵘 黄慧贤 CHEN Xinquan;ZHANG Zhan;ZHOU Weiguang;ZHANG Zhengrong;HUANG Huixian(Jiangmen customs district technology center,Jiangmen Guangdong 529000;Yangjiang customs district technology center,Yangjiang Guangdong 529500;Shantou customs district technology center,Shantou Guangdong 515000)

机构地区：[1]江门海关技术中心,广东江门529000 [2]阳江海关,广东阳江529500 [3]汕头海关技术中心,广东汕头515000

出　　处：《广东畜牧兽医科技》2024年第5期56-60,69,共6页Guangdong Journal of Animal and Veterinary Science

基　　金：海关总署科研项目(2021HK172);江门市基础与理论科学研究类科技计划项目(2020030100670005206)。

摘　　要：为制备猪塞内卡病毒(SVA)和O型口蹄疫病毒(FMDV-O)阳性质控品,根据选择的目的基因片段,设计阳性参考品序列,通过基因拼接技术和装甲RNA技术,制备含有口蹄疫O型病毒和塞内卡病毒目的基因的装甲RNA病毒样颗粒,用来作为塞内卡病毒(SVA)和O型口蹄疫病毒(FMDV-O)的阳性参考品。结果表明,该阳性质控品无生物传染性,可以真实模拟病毒粒子结构,耐受DNaseI及RNaseA,稳定性高,在-70℃保存18个月,荧光RT-PCR检测结果均无显著差异。本试验制备的含有猪O型口蹄疫病毒和塞内卡病毒目的基因的装甲RNA病毒样颗粒,可以作为SVA和FMDV-O双重荧光RT-PCR检测方法中进行质量控制的阳性参考品。To construct positive control samples for Seneca Valley virus(SVA)and foot-and-mouth disease virus type O(FMDV-O),positive reference sequences were designed based on selected target gene fragments.Armored RNA viral particles containing the target genes of FMDV-O and SVA were prepared through gene splicing and armored RNA technology.These particles were utilized as positive reference materials for SVA and FMDV-O.The results demonstrated that these positive control samples were non-infectious,faithfully simulated virus particle structures,were resistant to DNaseI and RNaseA degradation,exhibited high stability,and could be stored at-70℃for 18 months without significant changes in fluorescent reverse transcription polymerase chain reaction(RT-PCR)results.The prepared armored RNA viral particles containing the target genes of FMDV-O and SVA can be served as positive reference materials for quality control in the establishment of fluorescent RT-PCR detection methods for SVA and FMDV-O.

关 键 词：猪塞内卡病毒 O型口蹄疫病毒 MS2噬菌体 装甲RNA技术 阳性参考品 

分 类 号：S852.65[农业科学—基础兽医学]