摘要
为了快速检测单增李斯特菌,以李氏溶血素基因(hlyA)为靶基因,罗丹明B衍生物作为反应指示剂(阴性为红色,阳性为蓝色),通过优化指示剂、镁离子等浓度,建立一种快速、准确的环介导等温核酸扩增(LAMP)检测体系。结果表明:试剂盒法提取人工污染乳中该菌的基因组,LAMP体系扩增60min,通过肉眼观察扩增结果,检测限为1.08×101cfu/mL,其灵敏度是PCR法的10倍,与GB4789.30—2010相比,准确度为100%。
In order to detect Listeria monocytogenes, I choosed the hemolysin gene （hlyA） as the target gene, rhodamine-based dual chemosensor as the reaction indicator （negative in red, positive in blue）. At last, a fast, accurate loop-mediated isothermal nucleic acid amplification （LAMP） detection system was established by optimizing the indicator, magnesium ion concentration etc. The result indicated that the genome of bacteria, isolated from artificially contaminated milk, using the ‘‘Bacterial Genomic DNA Purification Kit’’, was amplified by the LAMP system for 60 min and the detection limit of L. monocytogenes was 1.08×10 1 cfu/mL with naked eyes. It’s sensitivity was 10 times than the PCR method, giving 100% concordance with the GB of 4789.30 —2010 reference method.
出处
《食品科技》
CAS
北大核心
2013年第1期322-326,共5页
Food Science and Technology
