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SPF级实验小鼠4种致病菌多重PCR方法的建立及优化
Authors
张越华
郑秀青
Publication date
28 August 2018
Publisher
Abstract
目的建立一种可同时检测4种SPF级屏障环境常见致病菌的多重PCR方法。方法本研究针对鼠伤寒沙门氏菌inv A基因、肺炎克雷伯杆菌khe基因、嗜肺巴斯德菌16SrRNA基因、铜绿假单胞菌ecf X基因序列分别设计合成特异性引物,构建可同时鉴别4种菌的多重PCR反应体系,优化后测定其特异性、灵敏性并人工模拟感染样本。结果建立的4重PCR方法能对同一样品中的4种致病菌模板进行特异性扩增,无交叉反应;对4种目的菌的检测灵敏度为10-3ng/μL;也可从混合感染的病料中特异性地检测出4种致病菌。结论本试验建立的多重PCR方法具有快速、简便、灵敏的特点,为4种致病菌的快速诊断和监控提供了有效的技术支持。福建省科技计划重点项目(No.2014Y0083
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Last time updated on 10/06/2020