Establishment of a method for detection of Mycobacterium abscessus by multiple cross-displacement amplification combined with hydroxynaphthol blue
YIN Shu-peng1, QIAN Cheng2, YAN Chen-qi3, LI Ma-chao3, LIU Hai-can3, LI Xiang-xin1, WAN Kang-lin3
1.Foshan Maternal and Child Health Care Hospital, Southern Medical University, Foshan 528000, China; 2. Beijing Center for Diseases Prevention and Control, Beijing 100013, China; 3. 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
Abstract: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 1.7×103 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.
尹树鹏, 钱城, 言晨绮, 李马超, 刘海灿, 李相新, 万康林. 多重交叉置换扩增联合羟基萘酚蓝检测脓肿分枝杆菌方法的建立[J]. 中国人兽共患病学报, 2020, 36(1): 40-44.
YIN Shu-peng, QIAN Cheng, YAN Chen-qi, LI Ma-chao, LIU Hai-can, LI Xiang-xin, WAN Kang-lin. Establishment of a method for detection of Mycobacterium abscessus by multiple cross-displacement amplification combined with hydroxynaphthol blue. Chinese Journal of Zoonoses, 2020, 36(1): 40-44.
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