中国兽医杂志2025，Vol.61Issue(3)：44-49,6.DOI:10.20157/j.cnki.zgsyzz.2025.03.006
猫传染性腹膜炎病毒SYBR Green实时荧光定量PCR检测方法的建立与临床应用
Establishment and Clinical Application of a SYBR Green Real-Time Fluorescence Quantitative PCR Detection Method for Feline Infectious Peritonitis Virus
摘要
Abstract
In order to improve the clinical detection rate of feline infectious peritonitis(FIP),this study designed specific primers targeting the M gene of feline infectious peritonitis virus(FIPV),cloned the amplified target gene into the pMD19-T vector,and obtained recombinant plasmids to be used as standard positive templates.A SYBR Green real-time fluorescence quantitative PCR(qPCR)detection method targeting FIPV was then established,and its specificity,sensitivity,reproducibility,and clinical detection rate were validated.The results showed that the standard curve of serially diluted standards exhibited a good linear relationship with Ct values(R2=0.999 9),and the amplification efficiency was 92.4%.There was no cross-reaction with other common feline viruses.The lowest detection limit was 3.48×102 copies/μL,which is at least 100 times more sensitive than conventional PCR methods.The method demonstrated excellent reproducibility,with intra-and inter-assay coefficient of variation both less than 2.4%.In clinical application,the detection rate for positive samples was 100%,while the detection rate for negative samples was 0%.The results indicate that the established SYBR Green qPCR method is specific,highly sensitive,reproducible,and effective for clinical FIPV diagnosis.关键词
猫传染性腹膜炎病毒/M基因/SYBR Green/实时荧光定量PCR(qPCR)Key words
feline infectious peritonitis virus/M gene/SYBR Green/real-time fluorescence quantitative PCR(qPCR)分类
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李星颖,白银山,陈胜锋,陈志胜,马骏,王丙云,谢梓民,原耀贤,孙铭澮,江文康,赵明明,何诗,何静,赖健仪..猫传染性腹膜炎病毒SYBR Green实时荧光定量PCR检测方法的建立与临床应用[J].中国兽医杂志,2025,61(3):44-49,6.基金项目
广东省普通高校动物干细胞工程技术研究中心项目(2021GCZX006) （2021GCZX006）
