Analytical performance of a multi-target open real-time PCR assay for simultaneous detection of tuberculosis, non-tuberculous mycobacteria, and drug resistance in a high-burden setting.
Microbiology spectrum·
- DOI
- 10.1128/spectrum.01643-26
- PMID
- 42603138
- PMCID
- —
- OpenAlex
- W7155552823
- Study type
- Journal article
- Publisher
- Publisher unavailable
- Article type
- journal-article
- Integrity
- current
Why this research matters now
Multi-target molecular platforms that detect both rifampicin and isoniazid resistance address current diagnostic gaps where rifampicin serves as the sole surrogate for multidrug resistance. The open-platform design may offer flexibility and reduced dependence on proprietary systems in high-burden settings.
Structured evidence summary
Research question
The study evaluated the analytical performance of two multiplex real-time PCR assays for concurrent detection of Mycobacterium tuberculosis complex, non-tuberculous mycobacteria, and resistance to rifampicin and isoniazid.
Study design
Analytical validation study using 300 well-characterized samples (150 MTBc-positive, 150 MTBc-negative) with composite reference standards including culture, MPT64 antigen testing, line probe assays, and phenotypic drug susceptibility testing.
Population and setting
The study was conducted in a high-burden tuberculosis setting using characterized culture isolates.
Main findings
MTBc detection achieved 100% sensitivity and specificity. NTM detection showed 70.0% sensitivity and 100% specificity. Rifampicin resistance detection demonstrated 96.0% sensitivity and 100% specificity, while isoniazid resistance detection achieved 100% for both measures. Agreement with reference standards was high (κ = 0.76-1.00).
Public-health relevance
Multi-target molecular platforms that detect both rifampicin and isoniazid resistance address current diagnostic gaps where rifampicin serves as the sole surrogate for multidrug resistance. The open-platform design may offer flexibility and reduced dependence on proprietary systems in high-burden settings.
Important limitations
NTM species coverage was incomplete. The study was an analytical validation using culture isolates rather than clinical specimens. Clinical validation, cost-effectiveness analysis, and operational feasibility studies have not yet been conducted.
GIDS interpretation
The article was identified through classifier links for tuberculosis diagnostics and treatment topics. It describes a laboratory validation study of a diagnostic technology rather than epidemiologic surveillance data.
Related GIDS surveillance
Literature context does not validate, explain, or change a surveillance signal. Exact and contextual relationships are shown separately.
Evidence relationships
This article has 7 auditable classifier relationships to diseases, places, topics, and study design.