细菌性感染性腹泻常见病原菌的多重PCR检测

目的:建立一种能同时检测细菌性感染性腹泻标本中沙门菌、志贺菌、金黄色葡萄球菌和副溶血性弧菌的多重聚合酶链反应(PCR)检测方法。方法:根据沙门菌的invA基因序列、志贺菌的ipaH基因序列、金黄色葡萄球菌的nuc基因序列、副溶血性弧菌的tdh基因序列设计特异性引物,通过多重PCR反应对样品中上述4种病原茵的目标基因进行扩增,同时优化反应体系。结果:本方法的特异性和灵敏性良好,该方法能检测的灵敏度分别为103.56pg的沙门菌DNA,94.85pg的志贺菌DNA,14.1pg的金黄色葡萄球菌DNA,115.32pg的副溶血性弧菌DNA。结论:该方法特异性和灵敏度高,检测周期短,适用于细菌性感染性腹泻标本中多种致病菌的快速检测,具有较好的应用前景。

Detection of pathogenic bacterium in infectious diarrhea by multiplex POR

Objective： To establish a multiplex PCR method for simultaneous detection of Salmonella spp. , Shigella spp. , Staphylococcus aureus, Vibrio parahaemolyticus in bacterial infectious diarrhea. Methods. The primers were designed according to gene invA of Salmonella spp. , gene ipaH of Shigella spp. , gene nue of Staphylococcus aureus and gene tdh of Vibrio parahaemolyticus. The target genes of these pathogens in bacterial infectious diarrhea were amplified by multiplex PCR, which was optimized specifically. Results： The multiplex PCR method established in this experiment was specific and accurate for detection of Salmonella spp （103.56pg）, Shigella spp. （94.85pg）, Staphylococcus aureus （14. 1pg） and Vibrio parahaemolyticus （115.32pg） respectively. Conclugiong： The multiplex PCR system is a quick and specific detection for bacterial infectious diarrhea, thus has great potential for clinical application in the future.