Combined Locked Nucleic Acid Probes and High-Resolution Melting Curve Analysis for Detection of Rifampicin-Resistant Tuberculosis in Northern Thailand
Issued Date
2022-10-01
Resource Type
eISSN
20754418
Scopus ID
2-s2.0-85140718508
Journal Title
Diagnostics
Volume
12
Issue
10
Rights Holder(s)
SCOPUS
Bibliographic Citation
Diagnostics Vol.12 No.10 (2022)
Suggested Citation
Thant Y.M., Saikaew S., Tharinjaroen C.S., Phunpae P., Pongsararuk R., Preechasuth K., Butr-Indr B., Intorasoot S., Tragoolpua K., Chaiprasert A., Wattananandkul U. Combined Locked Nucleic Acid Probes and High-Resolution Melting Curve Analysis for Detection of Rifampicin-Resistant Tuberculosis in Northern Thailand. Diagnostics Vol.12 No.10 (2022). doi:10.3390/diagnostics12102307 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/83584
Title
Combined Locked Nucleic Acid Probes and High-Resolution Melting Curve Analysis for Detection of Rifampicin-Resistant Tuberculosis in Northern Thailand
Author's Affiliation
Other Contributor(s)
Abstract
Rifampicin-resistant tuberculosis (RR-TB) has become a major threat globally. This study aims to develop a new assay, RIF-RDp, to enhance the detection of RR-TB based on combined locked nucleic acid (LNA) probes with high-resolution melting curve analysis (HRM). Two new LNA probes were designed to target the class-III and IV mutations of rpoB, H526D, and D516V. LNA probes showed 100% specificity in the detection of mutant targets among characterized and blinded Mycobacterium tuberculosis (Mtb) isolates. The performance of RIF-RDp was evaluated using 110 blinded clinical Mtb isolates in northern Thailand against drug-susceptibility testing (DST), DNA sequencing, and a commercial real-time PCR kit. This assay showed sensitivity and specificity of 94.55% and 98.18% compared to DST, and 96.36% and 100% compared to DNA sequencing. The efficacy of RIF-RDp was comparable to the commercial kit and DNA sequencing. The Cohen’s Kappa statistic showed almost perfect agreement between RIF-RDp and the commercial kit (κ = 0.95), and RIF-RDp and DNA sequencing (κ = 0.96). Furthermore, this is the first report of the rare mutation profiles, S531W, and a triple codon deletion (510–512) in northern Thailand. According to high accuracy, the RIF-RDp assay may render an easy-to-use, low-cost, and promising diagnostics of RR-TB in the future.