NDM-1基因新型染料EverGreen实时荧光PCR检测方法的建立

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

中文关键词:  NDM-1  EverGreen实时荧光PCR  检测  

英文关键词:NDM-1  EverGreen real-time fluorescent PCR  detection  

中文摘要:

      研究初步建立了NDM-1基因的实时荧光PCR快速检测鉴定方法。研究根据超级细菌NDM-1序列,设计并合成了检测引物(ND-E-F和ND-E-R),对NDM-1质粒DNA和其他11个标准菌株进行了EVERGREEN实时荧光PCR检测。结果表明,携有NDM-1基因的质粒DNA模板都出现了很强的特异信号,而其他对照病菌均未出现特异荧光信号,证明这套引物及探针具有NDM-1基因检测的特异性。灵敏度测定结果,此体系可检出10fg/μL质粒DNA模板。研究表明,EverGreen实时荧光PCR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 gene of superbacteria with broad-spectrum resistance to antibiotic.A EverGreen realtime PCR method was developed to detect NDM-1gene.Specific primers and EverGreen probe were designed based on the gene sequence in GeneBank.The probe 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 10 fg/μL DNA.Real-time fluorescent PCR is a specific,sensitive,rapid and highly effective method for the detection and identification of NDM-1.