摘要
本文建立了一个用于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.pneumophital-14、L.micdadei、L.dumoffii Llongbeachae、L.Jordanis、L.bozemanii),均可检出375bp的16SrRNA基因片段。对于已经监定的5株国内分离菌株染色体DNA进行扩增,获得了相同的结果,地高辛标记16SrRNA基因探针与扩增产物杂交,结果为阳性。而扩增14株非军团菌均为阴性。采用煮沸法制备细菌染色体DNA,PCR法检测环境水军团菌敏感性为280cfu/ml水,检查临床标本军团菌为560cfu/ml支气管灌洗液。上述结果表明该法敏感、快速、简便,具有较好的特异性。
A 16SrRNA polymerasse chain reaction(PCR)assay for the detection of Le-gionella Species was established.A 375 bp fragment of the 16SrRNA gene could be detected by amplifing genomic DNA of reference strains(Lp1-14,Lm,LI,Ld,Lb,Lg).T'he same result was also obtained by amplifing 5 identified strains isolated from China.The amplified products ware verified by hybridization with 16SrRNA gene probes.However,14 non-legionella strains could't produced a positive amplified fragment.Preparing bacterial genomic DNA by boiling method,the assay could detect 280cfu/ml water and 560 cfu/ml clinical bronchial fluid respectively.A bove-mentioned result showed that this assay was sensitive、rapid specific and easy to perform.
作者
宫仁梅
万超群
任红宇
卢锡华
Gong Shimei(Iastituie of Epid emiology and Microbiology,Chinese Acad emy of Preventive Medicimec,Beijing 102206)
出处
《疾病监测》
CAS
1996年第2期68-71,共4页
Disease Surveillance
