A new multiplex molecular assay based on padlock probes and rolling circle amplification for MDR-TB detection in clinical specimens: a prospective diagnostic accuracy study

一种基于锁式探针和滚环扩增技术的新型多重分子检测方法用于临床标本中耐多药结核病的检测:一项前瞻性诊断准确性研究

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Abstract

BACKGROUND: Patients with multidrug-resistant tuberculosis (MDR-TB) receive treatment that is at least 20% less effective than drug-susceptible cases globally. OBJECTIVES: The study evaluated the diagnostic accuracy of mfloDx MDR-TB assay (EMPE Diagnostics AB, Solna, Sweden) for simultaneous detection of resistance to isoniazid (INH) and rifampicin (RIF) compared to culture-based drug susceptibility test (DST). DESIGN: A prospective diagnostic accuracy study. METHODS: Clinical samples from 287 patients (mean age 45.3 (95% CI 43.3-47.2) years; 50 (17.0%) were female) from TB facilities in Ukraine (n = 82, 28.6%), Moldova (n = 37, 12.9%) and Spain (168, 58.5%) with bacteriologically confirmed TB and persons evaluated for non-tuberculous mycobacteria (NTM) were consecutively tested. The results of the mfloDx MDR-TB assay were compared with culture-based DST. RESULTS: A total of 186 out of 287 sputum specimens (64.8%) yielded conclusive results that allowed a definitive interpretation of resistance to at least one drug. The mfloDx MDR-TB assay demonstrated sensitivity at 86.9 (95% CI 80.6-91.7) and specificity at 100 (95% CI 79.4-100) correctly differentiating mycobacterium tuberculosis complex from NTM in smear positive sputum samples. Considering only conclusive results in sputum samples, the mfloDx MDR-TB assay simultaneously predicted resistance to INH and RIF in TB patients showing high sensitivity 100 (95% CI 93.0-100) and specificity 98.7 (95% CI 92.8-100) compared to MDR-TB detection by culture-based DST. CONCLUSION: While approximately one-third of tests yielded inconclusive results, the mfloDx™ MDR-TB assay demonstrated potential as a rapid screening tool for INH and RIF resistance, offering a time advantage over conventional culture-based DST.

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