广东畜牧兽医科技2024,Vol.49Issue(5) :56-60,69.DOI:10.19978/j.cnki.xmsy.2024.05.10

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

陈信全 张展 周伟光 张峥嵘 黄慧贤
广东畜牧兽医科技2024,Vol.49Issue(5) :56-60,69.DOI:10.19978/j.cnki.xmsy.2024.05.10

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

陈信全 1张展 1周伟光 2张峥嵘 3黄慧贤1
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作者信息

  • 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
  • 折叠

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.

Abstract

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

Key words

Seneca Valley virus/Foot-and-Mouth disease virus type O/MS2 bacteriophage/Armored RNA technology/Positive reference materials

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基金项目

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)

出版年

2024
广东畜牧兽医科技
广东省畜牧兽医学会 广东省农业科学院畜牧研究所 广东省农业科学院兽医研究所

广东畜牧兽医科技

影响因子:0.29
ISSN:1005-8567
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