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| Application of RPA-LFD for the rapid detection of rifampicin resistance in Mycobacterium tuberculosis |
| LU Yao1, ZHAO Li-li2, LI Ma-chao2, LIU Hai-can2, ZHAO Xiu-qin2, LIU Zhi-guang2, WAN Kang-lin2, LOU Yong-liang1 |
1. School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou 325035, China;
2. State Key Laboratory of Infectious Diseases Prevention and Control, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China |
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Abstract We established an RPA-LFD assay for the rapid detection of 531, 526 and 516 mutations in the Mycobacterium tuberculosis (MTB) rpoB gene, and evaluated the assay’s clinical application. We analyzed rifampicin resistance in 182 M. tuberculosis isolates preserved in the laboratory using drug sensitivity tests. Further analysis was performed by sequencing of the rifampin resistance-determining region (RRDR) in these strains and screening their mutation sites. The mutation sites in codons 516, 526 and 531 of rpoB were chosen and used to design primers for RPA-LFD. Finally, statistical analysis was conducted on the test results for the three methods to evaluate the detection efficiency of RPA-LFD. Relative to those of traditional DST, the sensitivity and specificity of RPA-LFD in the detection of rifampicin resistance were 84.2% and 100.0%, respectively. The Kappa value was 0.880, thus indicating high coincidence between the two methods. With respect to those of traditional DST, the sensitivity and specificity of DNA sequencing in the detection of rifampin resistance were 91.2% and 100.0%, respectively. RPA-LFD for the detection of resistance of M.tuberculosis to rifampicin showed good sensitivity and specificity, and was not time consuming; thus, it should have value in the rapid diagnosis of rifampicin-resistant tuberculosis.
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Received: 13 October 2020
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| Fund:Supported by the National Major Infectious Disease Prevention Projects(No.2018ZX10302302?001) |
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Corresponding Authors:
Lou Yong-liang, Email: louyongliang-2013@163.com
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2021, Vol. 37 
