A combination of Mycobacterium tuberculosis-derived sRNA and endogenous miRNA in circulation as novel auxiliary diagnostic markers for patients with multidrug-resistant tuberculosis.

OBJECTIVE

Circulating non-coding small RNAs (sRNAs) have recently emerged as promising biomarkers for Mycobacterium tuberculosis (Mtb). However, little is known about the expression patterns and combined diagnostic potential of M. tuberculosis-derived sRNA (TB-sRNA) and endogenous miRNA in the circulation of patients with multidrug-resistant tuberculosis (MDR-TB).

METHODS

Illumina sequencing by synthesis technology and quantitative real-time polymerase chain reaction were used to identify TB-sRNAs in serum and sputum from patients infected with drug-resistant TB, drug-sensitive TB (DS-TB), and controls. Simultaneously, four endogenous miRNAs - miR-29c, miR-132, miR-320b, and miR-548e - were chosen for further study. The diagnostic performance of significantly altered sRNAs and miRNAs was analysed using receiver operating characteristic (ROC) curves.

RESULTS

Illumina sequencing by synthesis combined with individual quantitative real-time polymerase chain reaction verification successfully identified one TB-encoded sRNA, named TB-sRNA015, and endogenous miR-132 were significantly elevated in patients with MDR-TB compared to patients with DS-TB and controls (P < 0.01). ROC analyses showed that the area under the ROC curve (AUC) for serum sRNA015 discriminating patients with MDR-TB from controls and patients with DS-TB were 0.774 (95% confidence interval [CI], 0.653-0.895) and 0.692 (95% CI, 0.560-0.825), respectively. For serum miR-132, the AUCs were 0.841 (95% CI, 0.735-0.946) for MDR-TB vs. controls and 0.655 (95% CI, 0.518-0.793) for MDR-TB vs. DS-TB. Importantly, combining sRNA015 with miR-132 increased the AUC to 0.860 (95% CI, 0.760-0.960) for discriminating MDR-TB from controls.

CONCLUSIONS

The combination of serum sRNA015 and miR-132 may serve as an auxiliary diagnostic tool for MDR-TB infection.