动物医学进展2025，Vol.46Issue(7)：11-17,7.
马疱疹病毒1型的RPA-CRISPR/Cas12a快速检测方法的建立
Establishment of RPA-CRISPR/Cas12a Rapid Detection Method for Equine Herpesvirus Type 1
摘要
Abstract
Equine herpesvirus 1(EHV-1)can cause equine rhinopneumonitis in equine animals,and its path-ological characteristics are mainly abortion in pregnant horses,respiratory infection and even death in new-born foals,which is a great harm to the equine industry.The aim of this study was to establish a rapid de-tection method of RPA-CRISPR/Cas12a for EHV-1 based on RPA amplification and CRISPR/Cas12a trans-digestion reaction of recombinant polymerase.During the experiment,similarity and genetic evolution of EHV-1-ORF68 gene were analyzed,and RPA primers and crRNA were designed according to their con-served sequences.RPA-CRISPR/Cas12a detection method of EHV-1 was established,and the specificity,sensitivity and clinical samples were tested.The results showed that the established RPA-CRISPR/Cas12a method of EHV-1 can detect EHV-1 specifically,and the detection of other equine-related pathogens was negative,with a high sensitivity and a minimum detection limit of 1.92 × 102 copies/μL.The sensitivity of this method was 100 times that of the traditional PCR method(1.92 × 104 copies/μL),and the detection time was short,with a total duration about 35 min.The established method was used to detect 24 clinical positive samples,and the positive detection rate of this method was(66.7％)higher than that of the tradi-tional PCR method(29.2％).In conclusion,the detection method of EHV-1 RPA-CRISPR/Cas12a estab-lished in this study has good specificity and sensitivity and accurate results,and can be used as an effective tool for rapid detection of EHV-1.关键词
马疱疹病毒1型/重组酶聚合酶扩增/CRISPR/Cas12a系统/快速检测Key words
Equine herpesvirus type 1/Recombinase polymerase amplification/CRISPR/Cas12a system/Rapid detection分类
农业科技引用本文复制引用
李银涛,撒瑞雪,宋毅,张嗣玉,刘建华,冉多良,邓海峰..马疱疹病毒1型的RPA-CRISPR/Cas12a快速检测方法的建立[J].动物医学进展,2025,46(7):11-17,7.基金项目
新疆维吾尔自治区天山英才"三农"骨干人才培养项目(2022SNGGGCC021) （2022SNGGGCC021）
新疆维吾尔自治区重大科技专项子课题(2022A02013-2-5) （2022A02013-2-5）
