摘要
目的建立一种可视化的恒温扩增方法用于检测脓肿分枝杆菌(MAB)。方法采用特异性全长erm(41)基因建立基于多交叉置换扩增(MCDA)结合羟基萘酚蓝(HNB)的方法,通过比色法直接检测扩增产物,采用23株脓肿分枝杆菌复合群和8种细菌标准株对该方法进行初步评价。结果建立的MAB-MCDA-HNB检测限为100 fg的DNA模板。31株细菌菌株中,只有脓肿分枝杆菌检测结果为阳性,其它细菌包括结核分枝杆菌,卡介苗,马赛分枝杆菌、鸟分枝杆菌等非结核分枝杆菌和金黄色葡萄球菌等均为阴性。脓肿分枝杆菌模拟痰标本检测灵敏度为1.7×10~3 CFU/mL(每次反应17 CFU)。结论MAB-MCDA-HNB方法能够快速准确诊断脓肿分枝杆菌的感染。
To establish a visualized isohermal amplification method for the detection of Mycobacterium abscessus(M. abscessus,MAB), a multiple cross displacement amplification(MCDA)-based assay combined with Hydroxynaphthol blue(HNB) was developed via specific detection of full-length erm(41) gene. The results showed that the absolute detection limit of MAB-MCDA-HNB was 100 fg DNA. Only M. abscessus was positive among the 31 strains of bacteria, while the other kinds including M. tuberculosis, BCG, Non-tuberculosis Mycobacterium(M. massilience, M.avium, etc) and Staphylococcus aureus were negative. Applied in the detection of M. abscessus in simulated sputum specimens, this method showed 1.7×10~3 CFU/mL(17 CFU each reaction) of the minimum detection concentration. The cost-effective and visible MAB-MCDA-HNB method developed here can diagnose M. abscessus infection quickly and accurately.
作者
尹树鹏
钱城
言晨绮
李马超
刘海灿
李相新
万康林
YIN Shu-peng;QIAN Cheng;YAN Chen-qi;LI Ma-chao;LIU Hai-can;LI Xiang-xin;WAN Kang-lin(Foshan Maternal and Child Health Care Hospital,Southern Medical University,Foshan 528000,China;Beijing Center for Diseases Prevention and Control,Beijing 100013,China;State Key Laboratory of Infectious Diseases Prevention and Control,Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases,National Institute for Communicable Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,China)
出处
《中国人兽共患病学报》
CAS
CSCD
北大核心
2020年第1期40-44,49,共6页
Chinese Journal of Zoonoses
基金
国家科技重大项目（No.2018ZX10302302-001-001,No.2018ZX10302302-001-005）资助~~
