PCR诊断细菌性痢疾的临床价值

目的建立一种快速、特异、敏感的诊断方法,对痢疾杆菌的致病基因进行基因检测研究。方法分别用PCR和多重聚合酶链反应(multiplex PCR)诊断方法,扩增2种痢疾杆菌的致病基因(iali、paH基因),扩增产物经电泳,出现与已知扩增片段大小一致的条带,并经测序与GENE库中标准菌株比较序列相同,即判断为该标本志贺菌的iali、paH基因阳性。结果对1组常规培养生化检测后的40份腹泻患者标本进行iali、paH基因特异片段扩增,ipaH基因阳性率为75%,ial基因阳性率为35%,iali、paH基因双基因阳性率为35%,较常规培养生化检测志贺菌阳性率(7.5%)高4倍多,iali、paH基因双基因阴性率为25%。结论PCR检测痢疾杆菌的致病基因具有简便、快速、特异、敏感和不需培养等特点,适于临床应用。

Development of the Multiplex PCR to Detect the Invasion Associated Genes of Shigella

Objective To establish a speed convenient and susceptive diagnosed method by detecting the invasion associated genes of shigella.Methods PCR and multiplex PCR was used to detect the pathogenic genes,ipaH and ial with the same time at one PCR reaction.Amplified DNA products of interest were sequenced,then the resulting was analyzed by use of the BLAST suite of programs.The sequences identified in the present databases available over the Internet were subsequently aligned.Results All 40 isolates from diarrhea stools were 14 positive for both ipaH and ial.Comparing with the routine methods the positive rate of the multiplex-PCR is high more than 4 times.Conclusion The method of PCR to detect the invasion associated genes of shigella is simple,special,sensitive,quickly and saving time.This new method can be widely used to clinic.