Impact of thermal pre-treatment on HDV RNA detection and quantification in clinical practice

Accurate detection and quantification of hepatitis delta virus (HDV) RNA are essential for diagnosis, prognosis and treatment monitoring. This study aimed to evaluate whether thermal shock pre-treatment improves HDV RNA detection and quantification using a commercial real-time RT-PCR assay. A total of 130 anti-HDV-positive serum samples were analysed with and without thermal shock pre-treatment prior to HDV RNA quantification. Qualitative and quantitative results were compared between untreated and heat-treated aliquots. Thermal shock increased viral load by ≥ 1 log in 78% of positive samples and enabled HDV RNA detection above the limit of detection in four samples initially classified as negative. These findings suggest that incorporating this step into routine workflows may reduce false negatives and represents a practical, low-cost, and impactful modification to current molecular protocols, directly strengthening clinical management and contributing to more reliable HDV surveillance.