中国兽药杂志2025，Vol.59Issue(5)：1-9,9.DOI:10.11751/ISSN.1002-1280.2025.5.01
鹅星状病毒Ⅰ、Ⅱ型和鹅细小病毒多重荧光定量PCR检测方法的建立与应用
Establishment and Application of Multiplex Quantitative PCR Methods for the Detection of Goose Astrovirus Type Ⅰ,Ⅱ and Goose Parvovirus
摘要
Abstract
To establish a rapid and specific multiplex real-time PCR method for the detection of Goose Astrovirus TypeⅠ(GAstVⅠ),Type Ⅱ(GAstVⅡ),and Goose Parvovirus(GPV),this study designed and synthesized specific primers and TaqMan probes with three different fluorescent labels based on the conserved gene sequences of GAstV Ⅰ,GAstV Ⅱ,and GPV.Recombinant plasmid standards containing the target genes of the three viruses were constructed.By screening primers,optimizing reaction conditions,and analyzing the specificity,sensitivity,and repeatability of the method,as well as the effect by detecting clinical samples,a multiplex real-time PCR detection method for GAstV Ⅰ,GAstV Ⅱ,and GPV was established.The results showed that this method has strong specificity,capable of specifically detecting GAstV Ⅰ,GAstV Ⅱ,and GPV without cross-reactivity with other common goose pathogens;high sensitivity,with a minimum detection limit of 10 copies/μL for the standard plasmids of GAstV Ⅰ,GAstV Ⅱ,and GPV;and good repeatability,with intra-and inter-assay coefficients of variation both less than 3％.When the detection results of 90 clinical samples using this method were compared with those obtained by previously published single real-time PCR methods for the three viruses.Among them,the coincidence rate of GAstV Ⅰ,GAstV Ⅱ and GPV was 98.89％、96.67％and 96.67％,respectively.The multiplex real-time PCR detection method established in this study can rapidly,specifically,and sensitively detect GAstV Ⅰ,GAstV Ⅱ,and GPV,enabling the detection of multiple pathogens in a single tube.It is highly practical and efficient,providing an effective technique for the monitoring and control of GAstV Ⅰ,GAstV Ⅱ,and GPV.关键词
鹅星状病毒Ⅰ型/鹅星状病毒Ⅱ型/鹅细小病毒/多重荧光定量PCRKey words
GAstV Ⅰ/GAstV Ⅱ/GPV/multiple real-time PCR分类
农业科技引用本文复制引用
朱啟超,任曼,张学刚,张进进,张鑫龙,李佳晋,曲光刚,李升和..鹅星状病毒Ⅰ、Ⅱ型和鹅细小病毒多重荧光定量PCR检测方法的建立与应用[J].中国兽药杂志,2025,59(5):1-9,9.基金项目
甘肃省科技重大专项计划(23ZDNA007) （23ZDNA007）
