中国动物检疫2024，Vol.41Issue(9)：105-110,6.DOI:10.3969/j.issn.1005-944X.2024.09.019
虾肝肠胞虫芯片式数字PCR定量检测方法的建立及应用
Establishment and Application of a cdPCR Assay for Enterocytozoon hepatopenaei
摘要
Abstract
Enterocytozoon hepatopenaei(EHP)infection has challenging global shrimp industry with a quick increasing prevalence over the past decade.In order to establish an absolute quantitative method for EHP,a set of specific primers and fluorescent probes were designed based on the conserved region of EHP SWP gene registered in GenBank,and a chip digital PCR(cdPCR)for EHP was established through optimizing reaction conditions.The results showed that the method had excellent linearity(R2=0.998 1);its detection limit was 4.4 copies/μL,indicating a high sensitivity;it failed to crossly react with other common shrimp pathogens,indicating a strong specificity;the variable coefficients of both the intra-and inter-group repeated tests were less than 8%,indicating a good repeatability.It was found that,for the detection of 28 clinical samples,the results by the established cdPCR method were 100%consistent with those by nested PCR proposed by the fishery industry standard of Enterocytozoon hepatopenaei disease Diagnosis Protocol(SC/T 7232-2020).In conclusion,the established method had good linear relationship,high sensitivity,strong specificity,good reproducibility,good accuracy and could achieve absolute quantification,indicating a good application prospect.关键词
虾肝肠胞虫/芯片式数字PCR/方法建立/性能评估Key words
EHP/cdPCR/method establishment/performance evaluation分类
农业科技引用本文复制引用
陈玉红..虾肝肠胞虫芯片式数字PCR定量检测方法的建立及应用[J].中国动物检疫,2024,41(9):105-110,6.基金项目
2024年福建省水生动物疫病监测与控制项目 （）
福建省海洋服务与渔业高质量发展专项资金项目(FJHY-YYKJ-2024-3-6) （FJHY-YYKJ-2024-3-6）
