Tuberculosis diagnosis and the complete drug resistance pattern from a single sample within a single day by use of a composite platform of MAX MDR-TB and AmPORE-TB

TL;DR

A new platform combines two tests to diagnose tuberculosis and determine drug resistance from a single sample in one day, improving treatment speed and outcomes.

Contribution

Introduces a one-day diagnostic platform integrating BD MAX MDR-TB and AmPORE-TB tNGS for rapid TB diagnosis and drug resistance profiling.

Findings

The platform achieved a final result in 8.5 hours with 85.1% sensitivity for spiked samples and 73% for clinical samples.

Resistance profiling failed most frequently for clofazimine, pretomanid, and delamanid in both spiked and clinical samples.

The platform reduces the need for multiple sputum samples and clinic visits, potentially lowering transmission and drop-out rates.

Abstract

Rapid TB diagnostics are essential for effective TB control. Combining WHO-recommended rapid molecular tests with downstream targeted next-generation sequencing (tNGS) enables faster drug resistance profiling. The objective of this study was to establish a one-day diagnostic platform (ODDP) integrating BD MAX MDR-TB and AmPORE-TB tNGS from a single sample. Pooled sputum samples spiked with 52 pre-characterized Mycobacterium tuberculosis (MTB) strains and 74 MTB-positive clinical samples were tested using BD MAX MDR-TB for TB, isoniazid, and rifampicin resistance. tNGS was performed from 5 µL of purified DNA leftover for each TB-positive sample in the BD MAX strips. IS6110/IS1081 Ct-values served as surrogate markers for TB DNA concentration. A total of 104 spiked and 60 clinical samples tested positive by BD MAX. The average time to the final ODDP result was 8.5 h. For samples with Ct ≤28, tNGS generated antibiotic resistance profiles for ≥12 antibiotics with 85.1% sensitivity in spiked and 73% in clinical samples. Failure rates were 10% and 8.3%, respectively. Resistance profiling most frequently (up to 11.3%) failed for clofazimine, pretomanid, and delamanid. The ODDP enables comprehensive TB diagnosis and resistance profiling from a single sample in 1 day. This platform can significantly accelerate the time to informed drug-resistant (DR)-TB treatment decisions.

Reducing the time to treatment initiation decreases patient drop-out rates, morbidity, the emergence of new drug resistances, and onward transmission of infection. Obtaining the complete resistome from the start is crucial for choosing a fully effective treatment regimen. Until now, diagnosis with full resistance profiling has required at least two sputum samples and 3 to 7 days for the complete workflow, obliging patients to return two to three times, which dramatically increased the risk of loss to follow-up. Our one-day diagnostic platform enables both diagnosis and comprehensive resistance testing from a single sample within 1 day. Patients can remain in a day clinic during testing and receive a fully effective, individualized treatment regimen the same day. This approach is expected to markedly reduce morbidity, drop-out rates, and transmission. The necessary instruments and technologies are already available in many high-prevalence countries and are currently being rapidly scaled up worldwide.

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