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．ｄｕｍｏｆｆｉｉＬｌｏｎｇｂｅａｃｈａｅ、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 165rRNA polymerasse chain reaction CPCR) 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, L1, Ld, Lb, Lg). The same result was also obtained by amplifing 5 identified strains isolated from China. The amplified products were 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. Above- mentioned result showed that this assay was sensitive、rapid、specific and easy to perform.
出处
《疾病监测》
CAS
1996年第2期68-71,共4页
Disease Surveillance
