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

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 .boze manni、L .dumoffii、L .longbeachae、L .micdadei、L .jordanis) ,均可检出 10 4bp的基因片段 ,敏感性为 8 5 2 10 2 cfu/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 10 3 cfu/ml。检测 17例血标本 ,阳性率为 5 8 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。

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*10 2cfu/ml(average)and 10 3cfu/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.