16s～23s rDNA间隔区序列聚合酶链反应鉴定术后龟分支杆菌暴发感染  

Identification of postoperative M chelonae infection outbreak by polymerase chain reaction of 16s～23s ribosomal DNA spacer sequence

作　　者：扈庆华[1] 李良成[1] 庄玉辉[2] 张灵霞[2] 刘军[1] 黄福新[1] 林和顺[1] 何建凡[1] 张顺祥[1] 

机构地区：[1]深圳市卫生防疫站,518020 [2]解放军第三○九医院

出　　处：《中华结核和呼吸杂志》1999年第11期648-651,共4页Chinese Journal of Tuberculosis and Respiratory Diseases

摘　　要：目的　从基因水平上确认深圳市某医院术后暴发感染的致病菌，并建立一套快速的ＰＣＲ方法鉴定龟分支杆菌脓肿亚种。方法　根据分支杆菌的１６ｓ～２３ｓｒＤＮＡ间隔区序列，设计合成一对引物和龟分支杆菌脓肿亚种ＤＮＡ探针，采用ＰＣＲ 和ＤＮＡ 斑点杂交技术，对５３ 株龟分支杆菌脓肿亚种临床分离株进行ＰＣＲ扩增和ＤＮＡ杂交。在此基础上，采用１６ｓ～２３ｓｒＤＮＡＰＣＲ扩增体系检测２５９份临床标本，对部分标本做ＤＮＡ探针斑点杂交反应。结果　５３ 株龟分支杆菌脓肿亚种临床分离株均被扩增出一条特异的３８０ ｂｐ ＤＮＡ 带和出现特异的杂交斑点。２５９ 份临床标本，ＰＣＲ 扩增阳性率为６０－６％ 。结论　在基因水平上确认此次引起院内术后暴发感染的致病菌为龟分支杆菌脓肿亚种。１６ｓ～２３ｓｒＤＮＡＰＣＲ扩增检测体系灵敏、特异，能鉴定龟分支杆菌。Objective To determine the pathogen of the infection outbreak in a hospital in Shenzhen at gene level with molecular biological technique, and to establish a rapid identification method of M chelonae subsp abscessus by polymerase chain reaction technique Methods A single pair of primers and oligonucleotides probe of M chelonae subsp abscessus were designed,according to the sequence of mycobacterium of 16s～23s ribosomal DNA spacer sequences 53 clinical strains were amplified by polymerase chain reaction, then dot blot hybridization of PCR product was made On the basis of this study, 259 leison, disease tissue and normal tissue were amplified by 16s～23s rDNA PCR Results 380 bp PCR product for 53 clinical strains was yielded by 16s～23s rDNA PCR, in the meantime, the specific hybridization dot appeared The PCR positive rate of 259 specimens was 60 6% Conclusions The PCR products and dot blot hybridization showed that the pathogen of the infection outbreak was M chelonae subsp abscessus at gene level The 16s～23s rDNA PCR investigative system is sensitive and specific, which can identify M chelonae subsp abscessus

关 键 词：外科伤口感染 龟分支杆菌 聚合酶链反应 

分 类 号：R63[医药卫生—外科学] R378.91[医药卫生—临床医学]