Further evidence of Mycobacterium tuberculosis in the sputum of culture negative pulmonary tuberculosis suspects using an ultrasensitive molecular assay

Abstract

Background: Rapid diagnosis of pulmonary tuberculosis (TB) is critical to TB control. However, many patients with paucibacillary TB disease remain undiagnosed. Current TB elimination goals require new tools to diagnose early disease. We evaluated performance of the Totally Optimized PCR (TOP) TB assay, a novel ultrasensitive molecular test. Methods: We assessed analytical specificity against nontuberculous mycobacteria (NTM), and estimated the diagnostic accuracy of TOP in a pilot study in Brazil (n = 46) and a cross-sectional study in Boston (n = 60). We compared TOP results to culture and a composite reference standard (CRS). Results: TOP exhibited no cross-reactivity against NTM. We tested 132 respiratory specimens from 106 patients with suspected pulmonary TB. The pilot demonstrated feasibility and 100% (95% CI 85–100) sensitivity in predominantly smear-positive specimens; TOP's specificity against solid media culture was low (58%, 37–77) but improved against a CRS (93%, 68–100). Similarly, when using the CRS in the Boston study, TOP (88%, 1–99) had greater sensitivity than solid or liquid media culture (25%, 3–65) and similar specificity (both 100%, 93–100). Conclusions: The TOP assay enables detection of M. tuberculosis in culture-negative paucibacillary disease. While the use of TOP for the diagnosis of paucibacillary disease will require further clinical validation, its high sensitivity indicate a more immediate utility as a rule out TB test.