环介导等温扩增技术在嗜水气单胞菌检测中的应用

发布时间：2018-05-27 02:24

本文选题：嗜水气单胞菌 + LAMP　；参考：《中国人兽共患病学报》2016年01期

【摘要】：目的建立环介导等温扩增技术(Loop-mediated isothermal amplification,LAMP)对嗜水气单胞菌检测的方法,并用临床标本对该方法进行评价。方法根据嗜水气单胞菌desA基因的保守序列设计特异性引物,利用参考质粒评价该检测体系的灵敏度;用30种其他肠道致病菌及院内感染中常见的致病菌评价该检测体系的特异性;用粪便模拟样本以及临床样本评价该检测体系的临床检测的可行性。结果 LAMP检测体系对嗜水气单胞菌重组质粒的检测灵敏度为1×102拷贝/反应体系;LAMP检测体系在检测30种其他肠道致病菌及院内感染中常见的致病菌时未出现假阳性;LAMP检测体系在粪便模拟标本中的检测下限为5×103 CFU/g;通过与qPCR技术相比较,LAMP具有更高的灵敏度以及更优秀的临床标本的检测能力。结论本研究建立的LAMP方法是首次将LAMP技术运用到了嗜水气单胞菌的检测,所需仪器设备简单,可作为一种快速的筛查方法在临床检验和基层实验室应用。
[Abstract]:Objective to establish a Loop-mediated isothermal amplification method for the detection of Aeromonas hydrophila and evaluate it with clinical specimens. Methods specific primers were designed according to the conserved sequence of desA gene of Aeromonas hydrophila and the sensitivity of the detection system was evaluated by reference plasmid. The specificity of the detection system was evaluated with 30 other intestinal pathogens and common pathogens in hospital infection, and the feasibility of clinical detection was evaluated with fecal simulated samples and clinical samples. Results the sensitivity of LAMP system for the detection of recombinant plasmids of Aeromonas hydrophila was 1 脳 10 ~ 2 copies / reaction system. The detection limit of lamp in fecal simulated samples is 5 脳 10 ~ 3 CFU / g. Compared with qPCR technique, lamp has higher sensitivity and better detection ability of clinical samples. Conclusion the LAMP method established in this study is the first time to apply the LAMP technique to the detection of Aeromonas hydrophila. The instrument and equipment required are simple and can be used as a rapid screening method for clinical examination and basic laboratory.
【作者单位】：中国疾病预防控制中心传染病预防控制所传染病预防控制国家重点实验室;
【基金】：国家重大传染病防治科技重大专项(2013ZX10004-101)资助~~
【分类号】：R446.5

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