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Lookup NU author(s): Dr Long Xie
This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).
© Springer-Verlag GmbH Germany, part of Springer Nature 2024.Background: Escalating cases of multidrug-resistant tuberculosis (MDR-TB) pose a major challenge to global TB control efforts, necessitating innovative diagnostics to empower decentralized detection of gene mutations associated with resistance to rifampicin (RIF) and isoniazid (INH) in Mycobacterium tuberculosis (M. tuberculosis) in resource-constrained settings. Methods: Combining multiplex fluorescent PCR and Multiple Probes Melting Analysis, we identified mutations in the rpoB, katG, ahpC and inhA genes from sputum specimens. We first constructed a reference plasmid library comprising 40 prevalent mutations in the target genes’ resistance determining regions and promoters, serving as positive controls. Our assay utilizes a four-tube asymmetric PCR method with specifically designed molecular beacon probes, enabling simultaneous detection of all 40 mutations. We evaluated the assay’s effectiveness using DNA isolated from 50 clinically confirmed M. tuberculosis sputum specimens, comparing our results with those obtained from Sanger sequencing and retrospective validation involving bacteriological culture and phenotypic drug susceptibility testing (pDST). We also included the commercial Xpert MTB/RIF assay for accuracy comparison. Results: Our data demonstrated remarkable sensitivity in detecting resistance to RIF and INH, achieving values of 93.33% and 95.24%, respectively, with a specificity of 100%. The concordance between our assay and pDST was 98.00%. Furthermore, the accuracy of our assay was comparable to both Sanger sequencing and the Xpert assay. Importantly, our assay boasts a 4.2-h turnaround time and costs only $10 per test, making it an optimal choice for peripheral healthcare settings. Conclusion: These findings highlight our assay’s potential as a promising tool for rapidly, accurately, and affordably detecting MDR-TB.
Author(s): Xie L, Zhu X-Y, Xu L, Xu X-X, Ruan Z-F, Huang M-X, Chen L, Jiang X-W
Publication type: Article
Publication status: Published
Journal: Infection
Year: 2024
Pages: epub ahead of print
Online publication date: 17/06/2024
Acceptance date: 10/05/2024
Date deposited: 24/06/2024
ISSN (print): 0300-8126
ISSN (electronic): 1439-0973
Publisher: Springer Science and Business Media Deutschland GmbH
URL: https://doi.org/10.1007/s15010-024-02295-w
DOI: 10.1007/s15010-024-02295-w
Data Access Statement: The manuscript and its supporting information fles encompass all pertinent data. Raw experimental data, including export fles of PCR reactions, and derived data supporting the fndings of this study can be available from the corresponding author X. Jiang upon reasonable request
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