Molecular Techniques for MTBC and NTM Differentiation: Diagnostic Accuracy of STANDARD™ M10 MTB/NTM and Potential Applications

Background. Over the past decade, the World Health Organization has highlighted the need for rapid molecular diagnostics as first-line tools for detecting Mycobacterium tuberculosis complex (MTBC) to strengthen global tuberculosis control. At the same time, infections caused by non-tuberculous mycobacteria (NTM) have become increasingly prevalent, particularly in low TB-burden countries such as Italy. This changing epidemiological scenario underscores the necessity for fast and reliable methods capable of distinguishing NTM from MTBC, a critical step for guiding appropriate treatment. This study evaluated the diagnostic accuracy and potential applications of the STANDARD™ M10 MTB/NTM assay, which simultaneously detects and differentiates MTBC and NTM. Methods. A total of 155 clinical specimens (78.1% respiratory) from patients with suspected mycobacterial infection were tested by fluorescence microscopy, GeneXpert MTB/RIF Ultra (respiratory samples only), STANDARD™ M10 MTB/NTM and culture, used as the reference method. Results. Culture detected MTBC in 54% and NTM (predominantly slow-growing species) in 46% of samples. STANDARD™ M10 showed overall sensitivity and specificity of 70% and 100%, respectively. For MTBC, sensitivity was 85.1% with almost perfect agreement with culture (κ = 0.866), while for NTM, sensitivity was 50% with moderate agreement (κ = 0.566). Sensitivity decreased in microscopy-negative/culture-positive specimens, particularly for NTM. Compared with GeneXpert MTB/RIF Ultra, STANDARD™ M10 exhibited slightly lower sensitivity for MTBC but retained excellent specificity. Conclusions. STANDARD™ M10 MTB/NTM represents a rapid, fully automated tool to support early etiological diagnosis and MTB/NTM differentiation, mainly in selected samples or high-risk patients, but it does not replace culture or molecular tests providing species identification and MTBC drug-resistance profiling.