J Luo1, M Luo2, J Li3, J Yu3, H Yang3, X Yi2, Y Chen2, H Wei3. 1. Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, University of Chinese Academy of Sciences, Beijing. 2. Chongqing Public Health Medical Center, Chongqing, China. 3. Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan.
Abstract
SETTING: Early diagnosis and drug susceptibility testing are important for anti-tuberculosis treatment. OBJECTIVE: To develop a rapid method for detecting Mycobacterium tuberculosis and drug susceptibility based on culture and droplet digital polymerase chain reaction (ddPCR). DESIGN: M. tuberculosis in 102 sputum samples was detected using ddPCR, Xpert, quantitative PCR (qPCR) and MGIT™ 960™. The susceptibility of ddPCR-positive samples to rifampicin (RMP), isoniazid (INH) and streptomycin (SM) was tested by measuring changes in DNA quantity over 4 days of culture. For comparison, susceptibility of MGIT 960-positive samples was tested using the standard agar proportion method. RESULTS: The sensitivity and specificity of M. tuberculosis detection using ddPCR and MGIT 960 were respectively 95.7% (95%CI 80.0-99.2) and 88.9% (95%CI 76.7-95.4). Compared with agar proportion, the susceptibility of 44 specimens positive on culture-ddPCR showed sensitivity and specificity for RMP, INH and SM of respectively 83.3% (95%CI 50.9-97.1) and 90.6% (95%CI 73.8-97.6); 79.0% (95%CI 53.9-93.0) and 92% (95%CI 72.5-98.6); 94.1% (95%CI 69.2-99.7) and 92.6% (95%CI 74.3-98.7). CONCLUSION: Culture ddPCR could detect M. tuberculosis within 5 h and drug susceptibility within 4 days directly from sputum, which would greatly reduce the laboratory time needed for tuberculosis diagnosis.
SETTING: Early diagnosis and drug susceptibility testing are important for anti-tuberculosis treatment. OBJECTIVE: To develop a rapid method for detecting Mycobacterium tuberculosis and drug susceptibility based on culture and droplet digital polymerase chain reaction (ddPCR). DESIGN:M. tuberculosis in 102 sputum samples was detected using ddPCR, Xpert, quantitative PCR (qPCR) and MGIT™ 960™. The susceptibility of ddPCR-positive samples to rifampicin (RMP), isoniazid (INH) and streptomycin (SM) was tested by measuring changes in DNA quantity over 4 days of culture. For comparison, susceptibility of MGIT 960-positive samples was tested using the standard agar proportion method. RESULTS: The sensitivity and specificity of M. tuberculosis detection using ddPCR and MGIT 960 were respectively 95.7% (95%CI 80.0-99.2) and 88.9% (95%CI 76.7-95.4). Compared with agar proportion, the susceptibility of 44 specimens positive on culture-ddPCR showed sensitivity and specificity for RMP, INH and SM of respectively 83.3% (95%CI 50.9-97.1) and 90.6% (95%CI 73.8-97.6); 79.0% (95%CI 53.9-93.0) and 92% (95%CI 72.5-98.6); 94.1% (95%CI 69.2-99.7) and 92.6% (95%CI 74.3-98.7). CONCLUSION: Culture ddPCR could detect M. tuberculosis within 5 h and drug susceptibility within 4 days directly from sputum, which would greatly reduce the laboratory time needed for tuberculosis diagnosis.
Authors: Adrienne E Shapiro; Jennifer M Ross; Mandy Yao; Ian Schiller; Mikashmi Kohli; Nandini Dendukuri; Karen R Steingart; David J Horne Journal: Cochrane Database Syst Rev Date: 2021-03-23
Authors: Jie Yi; Nan Wang; Jie Wu; Yueming Tang; Jingjia Zhang; Lingxiang Zhu; Xiao Rui; Yong Guo; Yingchun Xu Journal: Front Med (Lausanne) Date: 2021-12-22