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Rapid diagnosis of tuberculosis (TB) and antibiotic resistances are imperative to initiate effective treatment and to stop transmission of the disease. A new generation of more sensitive, automated molecular TB diagnostic tests has been recently launched giving microbiologists more choice between several assays with the potential to detect resistance markers for rifampicin and isoniazid. In this study, we determined analytical sensitivities as 95% limits of detection (LoD 95 ) for Xpert MTB/Rif Ultra (XP-Ultra) and BD-MAX MDR-TB (BD-MAX) as two representatives of the new test generation, in comparison to the conventional FluoroType MTB (FT-MTB). Test matrices used were physiological saline solution, human and a mucin-based artificial sputum (MUCAS) each spiked with  Mycobacterium tuberculosis  in declining culture- and qPCR-controlled concentrations. With BD-MAX, XP-Ultra, and FT-MTB, we measured LoD 95  TB  values of 2.1 cfu/ml (CI 95% : 0.9–23.3), 3.1 cfu/ml (CI 95% : 1.2–88.9), and 52.1 cfu/ml (CI 95% : 16.7–664.4) in human sputum; of 6.3 cfu/ml (CI 95% : 2.9–31.8), 1.5 cfu/ml (CI 95% : 0.7–5.0), and 30.4 cfu/ml (CI 95% : 17.4–60.7) in MUCAS; and of 2.3 cfu/ml (CI 95% : 1.1–12.0), 11.5 cfu/ml (CI 95% : 5.6–47.3), and 129.1 cfu/ml (CI 95% : 82.8–273.8) in saline solution, respectively. LoD 95  of resistance markers were 9 to 48 times higher compared to LoD 95  TB . BD-MAX and XP-Ultra have an equal and significantly increased analytical sensitivity compared to conventional tests. MUCAS resembled human sputum, while both yielded significantly different results than normal saline. MUCAS proved to be suitable for quality control of PCR assays for TB diagnostics.

plos one

A pre-clinical validation plan to evaluate analytical sensitivities of molecular diagnostics such as BD MAX MDR-TB, Xpert MTB/Rif Ultra and FluoroType MTB