NDM-1基因PCR检测技术的研究

The study of detection and identification of NDM-1 by PCR

NDM-1基因是一种"超级细菌"特有的耐药基因,对于菌体的广谱抗药性起着决定性的作用。本研究以实现该基因的快速检测为目的,初步建立了NDM-1基因的PCR快速检测鉴定法。选取肺炎克雷伯氏菌ADB65的NDM-1基因为目的基因,设计并合成了PCR检测引物(ND-319-F和ND-319-R),并以另外11种标准菌株为对照,分别进行了PCR检测。结果表明,携有NDM-1基因的质粒DNA经过PCR反应都扩增出长为319bp的特异性片段,而其他对照病菌均未出现相应片段,证明这对引物具有NDM-1基因检测的特异性。灵敏度测定结果,此体系可检出100fg/μL质粒DNA模板。研究表明,PCR方法是一种特异、灵敏、快速的NDM-1基因检测方法。

NDM-1 was an important gene of superbacteria with broad-spectrum resistance to antibiotic.A PCR method was developed to detect NDM-1gene.Specific primers were designed based on the gene sequence in GeneBank.The primers used in this assay were shown to be specific for plasmid with NDM-1 and did not react with other 11 standard bacteria strains. This technique was sensitive with the detection limit of 100 fg/μL DNA.PCR is a specific,sensitive,rapid and highly effective method for the detection and identification of NDM-1.