NDM-1基因DHPLC快速检测体系的建立

Arapid assay for detecting NDM-1 by DHPLC

中文关键词:  NDM-1  PCR-DHPLC  检测  

英文关键词:NDM-1  DHPLC  detection  

中文摘要:

      NDM-1基因是一种专属于"超级细菌"的重要特有的耐药基因,在食品潜在污染病原微生物的检测中着重要的作用。本研究以实现食品中超级细菌的快速检测为目标,初步建立了NDM-1基因的PCR-DHPLC快速检测鉴定方法,根据NDM-1基因序列,设计、合成了PCR引物(ND-D-F和ND-D-R),并分别对NDM-1质粒DNA及其他11个标准菌株进行了PCR扩增和DHPLC检测。结果表明,携有NDM-1基因的质粒DNA经过PCR-DHPLC反应扩增出特异性样品吸收峰,而其他对照菌均未出现相应吸收峰,提示本文中设计和应用的PCR引物具有NDM-1基因检测的特异性;灵敏度测定结果显示,此体系检测下限可达100fg/μL,提示PCR-DHPLC方法是一种特异、灵敏、快速的NDM-1基因检测方法。

英文摘要:

      NDM-1 was an important particular gene of superbacteria with broad-spectrum resistance to antibiotic,and make an important role in detecting pathogenic bacteria of food.For the aim at rapidly detect superbacteria in food,the method of detection and identification of NDM-1 by DHPLC was primarily established.A PCR-DHPLC method was developed to detect NDM-1 gene.Specific primers were designed and synthesized based on the gene sequence in GeneBank.The primers used in this assay were shown to be specific for plasmid with NDM-1 which produce special peak and other 11 standard bacteria strains. This technique was sensitive with the detection lower limit of 100 fg/μL DNA.PCR-DHPLC is a specific,sensitive,rapid and highly effective method for the detection and identification of NDM-1.