皮肤龟分枝杆菌感染的分子生物学鉴定  

Molecular biological identification on skin infection of Mycobacterium chelonae

作　　者：张忠奎[1] 康道现[2] 刘太华[1] 周舟[1] 王春梅[1] 王骏[1] 唐丽君[1] 陈业红[1] 冉玉平[2] 

机构地区：[1]中国人民解放军成都军区总医院皮肤科,成都610083 [2]四川大学华西医院皮肤性病科,成都610041

出　　处：《中国人兽共患病学报》2014年第11期1125-1128,共4页Chinese Journal of Zoonoses

摘　　要：目的用分子生物学方法鉴定龟分枝杆菌皮肤感染临床分离株。方法从1女性老年糖尿病患者右小腿化脓性感染灶,取创面分泌物沙堡琼脂培养基28℃培养4d,连续两次培养出光滑、湿润、不产色白色菌落。菌落涂片革兰染色为阳性杆菌,抗酸染色阳性。多次真菌镜检、培养阴性。皮损组织病理:皮下组织慢性化脓性感染,大量炎性细胞浸润,局部小脓肿形成;抗酸染色弱阳性杆菌,过碘酸雪夫(PAS)染色阴性。根据该菌株生长特征、生化反应,初步考虑为龟分枝杆菌,使用16SrRNA、rpoB、hsp65等分子生物学方法鉴定。结果 16S-rRNA引物测序结果于GenBank中做Blast比对,与龟分枝杆菌(Mycobacterium chelonae)AB548610.1同源性达100%,rpoB引物和hsp65引物测序结果做Blast比对与龟分枝杆菌同源性均为99%。结论该分离菌株为龟分枝杆菌。Molecular biological study was applied in identifying clinical isolated strain of Mycobacterium chelonae.The wound surface secretion was taken from the focus of suppurative infection in the right leg of a senile female patient with diabetes and was cultured in sabouraud＇s agar medium at the temperature of 28℃ for 4 days; the culture was performed for twice,smooth,wet and white nonphotochromogens colonies were gained,which were positive in Gram stain and acid-fast stain while negative in fungus microscope examination and culture.Histopathologic study of the skin lesion showed a chronic suppurative infection of subcutaneous tissue,inflammatory cell infiltration and local small abscess formation,while positive in acid-fast stain and negative in PAS stain.Molecular biological identification of 16S-rRNA,hsp65 and rpoB was performed and Blast intercomparasion was made in GenBank.The sequencing result of 16S-rRNA was 100％ in homology with Mycobacterium chelonae AB548610.1 while the sequencing results of rpoB and hsp65 were 99％ in homology with Mycobacterium chelonae AB548610.1.The strain is conformed to be Mycobacterium chelonae.

关 键 词：龟分枝杆菌 16S-rRNA HSP65 RPOB 

分 类 号：R378[医药卫生—病原生物学]