Abstract

Tuberculosis (TB) is an infectious disease caused by Mycobacterium tuberculosis and remains one of the leading causes of death worldwide. The increasing prevalence of TB strains resistant to multiple antibiotics, including isoniazid, highlights the urgent need for rapid and accurate molecular diagnostic methods. One such method is quantitative polymerase chain reaction (qPCR). This study aimed to compare the analytical sensitivity of qPCR assays for detecting isoniazid-resistant M. tuberculosis using SYBR Green and TaqMan Probe chemistries. Analytical sensitivity was evaluated using DNA from isoniazid-resistant M. tuberculosis carrying the S315G mutation, which was serially diluted up to 10⁻⁹. Primary data were analyzed using probit regression analysis. The results showed that the analytical sensitivity of the SYBR Green method was 1.5045 × 10⁻³ ng/µL, while that of the TaqMan Probe method was 1.0205 × 10⁻³ ng/µL. These findings indicate that the qPCR TaqMan Probe method is approximately 1.47 times more sensitive than the SYBR Green method. However, further studies evaluating additional validation parameters are required to determine the most suitable method for routine diagnostic application.

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