中国兽医科学2024，Vol.54Issue(9)：1182-1187,6.DOI:10.16656/j.issn.1673-4696.2024.0159
环形泰勒虫SHERLOCK-LF检测方法的建立与初步应用
Establishment and preliminary application of SHERLOCK-LF detection method for Theileria annulata
摘要
Abstract
To detect the infection of Theileria annulata quickly,accurately,and conveniently,T.annulata sporozoite surface antigen gene was selected as the target molecule to design specific RPA primers and crRNA(CRISPR RNA)sequences,and then a specific and highly sensitive enzyme reporter unlocking(SHERLOCK)-LF method was established to detect T.annulata infection.Meanwhile,the specificity,sensitivity and compliance of the method were analyzed.The results showed that the method could speci-fically detect the genome of T.annulata,and have no cross-reactivity with the genomes of Theileria sinen-sis,Theileria orientalis,Babesia bovis,or Babesia bigemina.As for its sensitivity,the method could detect 1.42 copies/μL and have 86.96％compliance compared with the PCR method.Furthermore the method could finish the detection of T.annulata in 1.5 h and provide technical support for its on-site diagno-sis,prevention and control.关键词
环形泰勒虫/RPA/Cas13a/crRNA/侧向流试纸条Key words
Theileria annulata/RPA/Cas13a/crRNA/lateral flow strip分类
农业科技引用本文复制引用
苏书晓,赵帅阳,刘军龙,张楚晗,朱浩瀚,关贵全,殷宏,罗建勋..环形泰勒虫SHERLOCK-LF检测方法的建立与初步应用[J].中国兽医科学,2024,54(9):1182-1187,6.基金项目
国家自然科学基金项目(32202836,31972706) （32202836,31972706）
国家重点研发计划项目(2022YFD1302102) （2022YFD1302102）
农业科技创新工程项目(CAAS-ASTIP-2016-LVRI) （CAAS-ASTIP-2016-LVRI）
国家肉牛牦牛产业技术体系项目(CARS-37) （CARS-37）
甘肃省基础研究创新群体项目(22JR5RA024) （22JR5RA024）
甘肃省科技厅特派团项目(22CX8NA011) （22CX8NA011）
国家寄生虫资源库项目(NPRC-2019-194-30) （NPRC-2019-194-30）
