Enhanced detection of rifampicin and isoniazid resistance in mycobacterium tuberculosis using AuNP-qPCR: a rapid and accurate method.

OBJECTIVES

To evaluate the resistance of Mycobacterium tuberculosis to Rifampicin (RIF) and Isoniazid (INH) using enhanced qPCR methodologies.

METHODS

This study compared the detection of drug-resistant mutations in the rpoB and katG genes using AuNP-qPCR and No-AuNP-qPCR. Calibration curves were constructed to correlate the amount of template with the Ct values for resistant strains.

RESULTS

The AuNP-qPCR method demonstrated high efficacy in detecting RIF resistance with an area under the curve (AUC) of 0.951, sensitivity of 97.92%, specificity of 87.5%, and overall accuracy of 95.31%. Similarly, INH resistance detection by AuNP-qPCR showed an AUC of 0.981, sensitivity of 98.08%, specificity of 94.44%, and accuracy of 97.14%. Comparatively, No-AuNP-qPCR yielded lower performance metrics for RIF resistance (AUC: 0.867, sensitivity: 91.67%, specificity: 75%, accuracy: 87.5%) and INH resistance (AUC: 0.882, sensitivity: 88.46%, specificity: 83.33%, accuracy: 87.14%).

CONCLUSIONS

AuNP-qPCR exhibits over traditional qPCR methods, making it a promising tool for rapid and precise detection of drug resistance in Mycobacterium tuberculosis. This method's robust performance underscores its potential to improve diagnostic protocols and contribute to more effective management of tuberculosis treatment.