ArticleFrontiers in microbiology2026
Analytical validation of a multiplex PCR assay for differentiation of five clinically significant nontuberculous mycobacterial species.
Article in Frontiers in microbiology, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Background: In TB endemic areas, accurate identification of non-tuberculous mycobacteria (NTM) from Objective: To develop and analytically validate an in-house multiplex PCR (mPCR) assay for simultaneous differentiation of five clinically relevant NTM species. Methods: The assay was developed using reference strains of Results: The in-house mPCR technique successfully detected all five target species simultaneously and showed clear species-specific amplification, with distinct amplicon patterns (178-392 bp). The analytical limit of detection was established at 1 ng/μL using purified DNA, with reproducible amplification across replicate reactions. Spiked sample evaluation confirmed detection capability in complex biological matrices, though reduced band clarity was observed at lower concentrations. High analytical specificity was confirmed by the absence of cross-reactivity with non-target species, such as Conclusion: The developed in-house mPCR assay offers a rapid, economical molecular method for differentiating clinically important NTM species and has strong analytical performance. To justify its use in standard diagnostic labs, further extensive clinical validation is necessary.
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