中国兽医杂志2025，Vol.61Issue(5)：43-51,9.DOI:10.20157/j.cnki.zgsyzz.2025.05.005
基于MS2噬菌体的小反刍兽疫装甲RNA质控品的研制
Preparation of MS2 Phage-based Armored RNA Quality Control for Peste des Petits Ruminants
摘要
Abstract
To develop a safe and stable quality control material for nucleic acid detection of peste des petits ruminants virus(PPRV),this study constructed a recombinant plasmid(pET-CPA-PPR)by inserting the target gene N of PPRV,the coat protein(CP)gene of MS2 phage,and the maturase protein A gene sequence into the pET28a vector.After expression,purification,removal of residual nucleic acids,and verification by real-time fluorescence quantitative PCR,the PPRV armored RNA quality control material was prepared.The quality control material was then evaluated by transmission electron microscopy,quantification,nuclease resistance,homogeneity,and stability testing to assess its potential as a certified reference material.The results showed that the pET-CPA-PPR construct,after induction and purification,produced uniformly sized armored RNA particles with diameters ranging from 23 to 28 nm,encapsulating the PPRV target gene N,with a concentration of 2.24×10⁹ copies/µL.These particles demonstrated strong resistance to nuclease degradation,good homogeneity,and remained stable for at least 30 days at(37±1)℃.In conclusion,the PPRV armored RNA prepared based on MS2 bacteriophage exhibited high copy number,excellent homogeneity and stability,and can serve as a safe and reliable standardized quality control material for nucleic acid detection of PPRV.关键词
小反刍兽疫(PPR)/MS2噬菌体/装甲RNA/病毒样颗粒Key words
peste des petits ruminants(PPR)/MS2 phage/armored RNA/virus-like particles分类
畜牧业引用本文复制引用
刘丹,熊涛,沈青春,李慧彤,张博源,高建帅,蒋卉,范学政,张广智,郜卫华,丁家波..基于MS2噬菌体的小反刍兽疫装甲RNA质控品的研制[J].中国兽医杂志,2025,61(5):43-51,9.基金项目
"十四五"国家重点研发计划项目-重大外来动物疫病阻断与防控技术研发项目(2022YFD1800500) （2022YFD1800500）
