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空肠弯曲菌环介导等温扩增检测方法的建立和优化

DOI: 10.3969/j.issn.1000-7091.2013.03.040, PP. 222-226

Keywords: 空肠弯曲菌,环介导等温扩增方法,检测,优化

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Abstract:

首先根据空肠弯曲菌mapA基因序列设计LAMP引物,建立检测空肠弯曲菌的LAMP检测方法。继而对LAMP方法中不同反应组分的浓度分别进行筛选,结果显示,当内外引物浓度分别为1.6,0.2μmol/L、Mg2+。浓度为8mmol/L、dNTP浓度为1.4mmol/L、Betaine浓度为0.8mol/L时获得最佳的反应结果。最后对LAMP反应的特异性和敏感性进行检测,特异性检测结果显示,对其他13种革兰氏阴性和革兰氏阳性细菌扩增结果均为阴性;敏感性检测结果显示,对空肠弯曲菌基因组DNA的检测限为每个反应管100fg,对空肠弯曲菌培养菌液检测限为每个反应管7.5cfu。试验建立的LAMP方法为快速简便地自动物源性产品中检测空肠弯曲菌奠定了基础。

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