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

Autoři: Markus Beutler aff001;  Sara Plesnik aff001;  Marina Mihalic aff001;  Laura Olbrich aff002;  Norbert Heinrich aff002;  Samuel Schumacher aff004;  Michael Lindner aff005;  Ina Koch aff005;  Wolfgang Grasse aff006;  Christoph Metzger-Boddien aff006;  Sabine Hofmann-Thiel aff001;  Harald Hoffmann aff001
Působiště autorů: Institute of Microbiology and Laboratory Medicine, Department IML Red GmbH, WHO-Supranational Reference Laboratory of Tuberculosis, Munich-Gauting, Germany aff001;  Division of Infectious Diseases and Tropical Medicine, LMU University Hospital, Munich, Germany aff002;  German Centre of Research on Infectious Diseases (DZIF), Partner Site Munich, Germany aff003;  Foundation for Innovative New Diagnostics, Geneva, Switzerland aff004;  Asklepios Fachkliniken München-Gauting, Munich, Germany aff005;  gerbion GmbH & Co. KG, Kornwestheim, Germany aff006;  SYNLAB Gauting, SYNLAB Human Genetics Munich, Munich-Gauting, Germany aff007
Vyšlo v časopise: PLoS ONE 15(1)
Kategorie: Research Article
doi: https://doi.org/10.1371/journal.pone.0227215


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 (LoD95) 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 LoD95TB values of 2.1 cfu/ml (CI95%: 0.9–23.3), 3.1 cfu/ml (CI95%: 1.2–88.9), and 52.1 cfu/ml (CI95%: 16.7–664.4) in human sputum; of 6.3 cfu/ml (CI95%: 2.9–31.8), 1.5 cfu/ml (CI95%: 0.7–5.0), and 30.4 cfu/ml (CI95%: 17.4–60.7) in MUCAS; and of 2.3 cfu/ml (CI95%: 1.1–12.0), 11.5 cfu/ml (CI95%: 5.6–47.3), and 129.1 cfu/ml (CI95%: 82.8–273.8) in saline solution, respectively. LoD95 of resistance markers were 9 to 48 times higher compared to LoD95TB. 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.

Klíčová slova:

Antibiotic resistance – Multi-drug-resistant tuberculosis – Mycobacterium tuberculosis – Polymerase chain reaction – Sputum – Tuberculosis – Tuberculosis diagnosis and management – Saline solutions


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