华南农业大学学报2025，Vol.46Issue(3)：311-318,8.DOI:10.7671/j.issn.1001-411X.202403028
基于PCR-CRISPR/Cas12a技术的肠炎沙门菌快速检测方法
A fast method for detecting Salmonella enteritidis based on PCR-CRISPR/Cas12a
摘要
Abstract
[Objective]To realize early detection,prevention and control of Salmonella enteritidis,a detection method was established by combining CRISPR(Clustered regularly interspaced short palindromic repeat)technology with polymerase chain reaction(PCR).[Method]PCR primers were screened based on the conserved sdfI gene sequence.The sensitivity test was conducted using S.enteritidis genomes at different concentrations,while the specificity tests were conducted using the genomes of Escherichia coli,Staphylococcus aureus,Pasteurella multocida,Salmonella choleraesuis and Enterococcus faecalis.Subsequently,the developed method was applied for the detection of S.enteritidis in small intestinal samples.[Result]This method was capable of identifying S.enteritidis in small intestinal samples,with a detection limit of 1.72×100 μL-1,showing excellent specificity and no cross-reactivity with E.coli,S.aureus,P.multocida,S.choleraesuis and E.faecalis.[Conclusion]This study develops a compact diagnostic method for the early detection of S.enteritidis,showing excellent sensitivity and specificity.This innovation presents a fresh perspective for the swift identification of S.enteritidis.关键词
肠炎沙门菌/sdfI基因/聚合酶链式反应/CRISPR/Cas12a系统Key words
Salmonella enteritidis/sdfI gene/PCR/CRISPR/Cas12a system分类
农业科技引用本文复制引用
杨天沐,王毅恒,熊文广,郭剑英..基于PCR-CRISPR/Cas12a技术的肠炎沙门菌快速检测方法[J].华南农业大学学报,2025,46(3):311-318,8.基金项目
国家重点研发计划(2022YFD1800400) （2022YFD1800400）
广东省基础与应用基础研究基金联合基金(2023A1515030137) （2023A1515030137）
