中国兽医科学2025，Vol.55Issue(3)：330-337,8.DOI:10.16656/j.issn.1673-4696.2025.0037
绵羊无浆体 LAMP-CRISPR/Cas12a检测方法的建立
Development of LAMP-CRISPR/Cas12a assay for the detection of Anaplasma ovis
摘要
Abstract
Anaplasma ovis is a tick-borne bacterial pathogen that parasitizes erythrocytes,caus-ing disease in sheep,goats,and wild ruminants.It poses a serious threat to the development of the sheep farming industry in China.Although various diagnostic methods for A.ovis have been reported,there is still a lack of simple and rapid detection techniques for field application.This study aimed to estab-lish a LAMP-CRISPR/Cas12a detection method targeting the msp4 gene of A.ovis.This detection method is conducted at a constant temperature and dose not require special equipment,allowing for the generation of fluorescent results that could be visually interpreted.By optimizing the system for the LAMP-CRISPR/Cas12a method,the optimal concentration ratios of the components were determined.The method could detect a minimum plasmid DNA copy number of 3.90 copies/μL for the msp4 gene of A.ovis,demonstrating high sensitivity.Additionally,the results were negative for other Anaplasma or common sheep pathogens,indicating good specificity.In clinical sample applications,the consistency with conventional PCR results was 100％.The results suggested that the established LAMP-CRISPR/Cas12a system is suitable for the detection of A.ovis,offering advantages for rapid and convenient field di-agnosis and application in resource-scarce situations.关键词
绵羊无浆体/msp4基因/LAMP/CRISPRKey words
Anaplasma ovis/msp4/LAMP/CRISPR分类
畜牧业引用本文复制引用
郭心龙,周勇志,曹杰,王亚楠,张厚双,段德勇,周金林..绵羊无浆体 LAMP-CRISPR/Cas12a检测方法的建立[J].中国兽医科学,2025,55(3):330-337,8.基金项目
国家重点研发计划项目(2022YFE0120100) （2022YFE0120100）
