CAJ | 학술논문

目的：建立一种环介导等温扩增（LAMP）反应快速检测大肠埃希菌的方法。方法根据美国国立卫生研究院基因序列数据库（Genbank）上大肠埃希菌的lacZ（登录号：M74750）基因序列，设计4条特异引物（2条内引物，2条外引物），对lacZ基因进行扩增，对扩增反应进行优化，采用琼脂糖电泳及肉眼观察结果。结果将提取的13株细菌DNA进行LAMP反应，仅大肠埃希菌得到阳性结果，LAMP检测下限为15 cfu/mL。结论 LAMP检测大肠埃希菌是快速、低成本、特异性强、灵敏度高的方法，适合临床开展。
목적：건립일충배개도등온확증（LAMP）반응쾌속검측대장애희균적방법。방법근거미국국립위생연구원기인서렬수거고（Genbank）상대장애희균적lacZ（등록호：M74750）기인서렬，설계4조특이인물（2조내인물，2조외인물），대lacZ기인진행확증，대확증반응진행우화，채용경지당전영급육안관찰결과。결과장제취적13주세균DNA진행LAMP반응，부대장애희균득도양성결과，LAMP검측하한위15 cfu/mL。결론 LAMP검측대장애희균시쾌속、저성본、특이성강、령민도고적방법，괄합림상개전。
Objective To apply a one-step loop-mediated isothermal amplification(LAMP) assay for rapid detection of Escherichiaco-li .Methods According to the Genbank of Escherichiacoli lacZ(accession number:M74750) ,a set of four specific primers were designed for the Escherichiacoli LAMP assay and optimized the reaction .The results were judged with naked eyes and electrophoretic analysis .Results Genomic DNAs from 13 bacterial strains including Escherichia coli strains were amplified using LAMP ,and no amplicon was observed in the other bacterial strains .The detection limits of LAMP assay was 15 cfu/mL .Conclusion LAMP assay is extremely rapid ,cost-effective , specific and highly sensitive and has potential usefulness for clinical application .