【作者】 王国戗； 罗予；

【Author】 WANG Guo-Qiang LUO Yu Basic medical college of zhengzhou university Henan institute of medical science

【机构】 郑州大学基础医学院； 河南省医学科学研究所；

【摘要】 类杆菌属细菌是一群G-厌氧杆菌,也是临床重要的条件致病菌,快速, 准确鉴定这类细菌对临床感染性疾病的诊断和治疗非常重要。但是,传统的鉴定方法费时费力,一般需要2-7天时间。目的:对该属细菌进行PCR鉴定,以缩短鉴定时间,为临床诊断及时提供依据。建立该属细菌的基因进化树,结合基因序列比较(Blast),以观察不同来源的该属细菌的变异情况。方法:采用16S rRNA基因序列为靶基因设计引物,对该属标准菌株及不同来源的临床株进行 PCR扩增,然后对扩增产物进行基因测序验证引物的特异性并建立基因进化树。结果:建立的该PCR反应体敏感性为100个细菌基因组,而40株肠杆菌科细菌、本实验室分离和保存的其他非类杆菌不能被扩增;建立的基因进化树表明不同来源的该属细菌16srRNA基因序列比较,同源性达98％。在该实验过程中我们发现了一个新的基因序列,该序列已经被GenBank数据库接收,接收号为 DQ646554。更多还原

【Abstract】 The Bacteroides are vital Bacteria that caused disease in clinic. So timely and exact identification of the Bacteroides is very important for diagnose and therapy of infectious diseases. However, conventional identified methods spend too many time and human power , and need 2-7 days to identify these bacteria .Objective: Quickly identify Bacteroides species by developing PCR assay . Methods:To design Primers based on 16srRNA gene sequences; measure sequences of PCR products ; To establish gene evolvement tree .through comparing of comparison of sequences by CLUSTAL W1.83 soft ware ,study the differentiation of bacteria which come from different origins . Results: The sensitivity is 100 gen group , and the method had very fine specificity. The established gene tree showed that 16s rRNA gene sequences of different origins are relatively conservative, the lowest homology come to98% between these sequences. We find a new gene sequence, which has been received by GenBank , remarked number is DQ646554 ,during our work.更多还原