摘要
目的:建立一种快速明确饮用水中是否存在细菌污染的核酸检测方法。方法:根据细菌16SrRNA中高度保守的序列设计4条LAMP引物,交由商业公司合成;利用合成好的引物和标准菌株,通过LAMP方法验证引物扩增的效率和特异性,电泳观察扩增结果,并比较两种不同细菌基因组DNA提取方法及优化LAMP反应条件。结果:使用设计的引物可以很好的检测出各种标准菌株,而且检测灵敏度达到要求。使用碱裂解的方法可以非常方便的提取出细菌基因组DNA。结论:成功建立了LAMP法检测饮用水中细菌的方法,为饮用水是否存在细菌污染快速检测提供了一种可用的方法。
Objective：To establish a nucleic acid detection method for rapidly testing bacteria in potable water. Methods： According to the high conserve sequence in bacteria 16sRNA gene, LAMP primers were designed and synthesized by a commercial company. Then the amplification efficiency and specificity were detected using synthesized primers and standard bacteria strains through LAMP, the arnplification result was observed through electrophoresis. Two kinds of bacteria genome preparing methods were also compared. Results： All standard bacteria strains could be detected using synthesized primers. The detection sensitivity could meet the requirement. Alkali cracking method was an easy and convenient mean to prepare bacteria genome. Conclusion： A method based on LAMP for detecting bacteria in drinking water was established successfully, lit gave us a rapid way to judge whether potable water was contaminated by bacteria.
出处
《中国卫生检验杂志》
北大核心
2013年第3期644-646,共3页
Chinese Journal of Health Laboratory Technology
