军团菌16SrRNA聚合酶链反应方法的建立及应用

Establishment and Application of a 16SrRNA Polymerase Chain Reaction Assay for Detection of Legionella Species

建立了１６ＳｒＲＮＡ基因检测军团菌的多聚酶链反应方法。扩增参考菌株染色体ＤＮＡ（Ｌ．Ｐｎｅｕｍｏｐｈｉｌａ１～１４、Ｌ．ｂｏｚｅｍａｎｎｉ、Ｌ．ｄｕｍｏｆｉ、Ｌ．ｌｏｎｇｂｅａｃｈａｅ、Ｌ．ｍｉｃｄａｄｅｉ、Ｌ．ｊｏｒｄａｎｉｓ），均可检出出３８６ｂｐ的基因片段，敏感性为１０３ｃｆｕ／ｍｌ（平均值）；而扩增９株非军团菌均为阴性。对模拟血标本的检测，其敏感性可达１０２ｃｆｕ／ｍｌ。应用本检测系统检测了２４例临床标本，包括１９份血、５份胸水，阳性率为７０．８％（１７／２４）。与血清学检测和临床诊断治疗结果相符合。上述结果表明该方法敏感、特异、快速、简便。

A 16SrRNA Polymerase Chain reaction(PCR)assay for the detection of Legionella Species was established.A 386bp fragment of the 16SrRNA gene could be detected by amplifing genomic DNA of reference strains(Lp1～14、Lb、Ld、Ll、Lm、L;).9 non legionella strains couldn't produced a positive amplified feagment.The sensitive for reference strains and blood were 10 3cfu/ml(average)and 10 2cfu/ml respectively.This PCR system has been successful in detecting 24 clinical samples,the positive rate was 70.8%(17/24).Above mentioned result showed that this assay was sensitive,rapid,spesific and easy to perform.