Improved heat shock method for extracting total RNA from nasopharyngeal swab samples even with low viral load

This study aimed to improve the heat shock method as a cost-effective and time -efficient for total RNA extraction. We compared the effectiveness of two total RNA extraction methods by using Real -Time PCR for nasopharynx swabs. Include: I; use of a commercial total RNA extraction kit as a standard. II; utilized a modified heat shock method (MHS). Time, centrifuge speed and duration, proteinase K, and RNA carrier were optimized. The optimized parameters included treating the sample with 5 mu g/mu L at 56(degrees)C for 5 minutes, heating at 95(degrees)C for 5 minutes followed by thermal shock in ice for 3 minutes, adding 4 mu g/mu L RNA carrier at room temperature for 3 minutes, and centrifuging at 7000 rpm for 10 minutes. This optimization demonstrated a sensitivity and specificity of 100% (CI: 95%) even in samples with low viral load. Our in-house method presents a rapid, and cost-effective alternative for total RNA extraction.