军团菌5SrRNA聚合酶链反应方法的建立及应用

Establishment and Application of a 5SrRNA Polymerase Chain Reaction Assay for Detection of Legionella Species

摘要: 建立5SrRNA基因检测军团菌的多聚酶链反应方法,扩增参考菌株的染色体DNA(L.Pneumophila1～14、L.bozemanni、L.dumoffii、L.longbeachae、L.micdadei、L.jordanis),均可检出104bp的基因片段,敏感性为8.52*10²cfu/ml(平均值);而扩增9株非军团菌均为阴性。对模似血标本的检测,其敏感性可达10³cfu/ml.检测17例血标本,阳性率为58.8%(10/17),与血清学检测和临床诊断治疗结果相符合。结果表明该方法敏感、特异、快速、简便。

Abstract: A 5SrRNA Polymerase Chain reaction(PCR)assays for the detection of Legionella Species was established.A 104bp fragment of the 5SrRNA gene could be detected by amplifing genetic DNA of reference strains(Lp1～14、Lb、Ld、Ll、Lm、Lj).9 non legionella strains could not produce a positive amplified fragment.The sensitive for reference strains and blood were 8.5*10²cfu/ml(average)and 10²cfu/ml respectively.This PCR system has been successful in detecting 17 clinical samples,the positive rate were 58.8% (10/17).Above mentioned result showed that this assay was sensitive,rapid,specific and easy to perform.