浙江农业学报2025，Vol.37Issue(12)：2468-2478,11.DOI:10.3969/j.issn.1004-1524.20240730
十足目虹彩病毒1和虾肝肠胞虫双重荧光定量PCR检测方法的建立
Establishment of a double quantitative PCR method for the detection of Decapod iridescent virus 1 and Enterocytozoon hepatopenaei
谈荣想 1斯广杰 2孙海涛 2许婷1
作者信息
- 1. 绍兴文理学院 生命与环境科学学院,浙江 绍兴 312000
- 2. 浙江华珍科技有限公司,浙江 诸暨 311804
- 折叠
摘要
Abstract
To establish a method for simultaneous detection of Decapod iridescent virus 1(DIV1)and Enterocytozoon hepatopenaei(EHP),this study developed a double SYBR Green Ⅰ-based quantitative real-time PCR(qPCR)as-say,building upon an established qPCR method for DIV 1.The assay targets the major capsid protein(MCP)gene of DIV1 and the spore wall protein(SWP)gene of EHP.The results showed that the melting temperatures(Tm)for DIV1 and EHP were(82.0±0.5)℃and(77.0±0.5)℃,respectively.The method specifically detected DIV1 and EHP,while showing negative results for other common shrimp pathogens and healthy shrimp samples.Within 35 amplification cycles,the limits of detection were 75 copies·μL-1 for DIV1 and 15 copies·μL-1 for EHP.Reproduc-ibility tests showed that both intra-and inter-assay coefficients of variation were below 2%.In conclusion,the estab-lished double qPCR method exhibits high specificity,sensitivity,and reproducibility,and is suitable for rapid moni-toring of DIV1 and EHP infections in shrimp.关键词
双重SYBR Green Ⅰ实时荧光定量PCR/十足目虹彩病毒1/虾肝肠胞虫/虾Key words
double SYBR Green Ⅰ quantitative real-time PCR/Decapod iridescent virus 1/Enterocytozoon hepatope-naei/shrimp分类
农业科技引用本文复制引用
谈荣想,斯广杰,孙海涛,许婷..十足目虹彩病毒1和虾肝肠胞虫双重荧光定量PCR检测方法的建立[J].浙江农业学报,2025,37(12):2468-2478,11.
