Clinical Performance Characteristics of Hepatitis B Virus Quantification by Combining Artus-DSP Assay with Rotor Gene Q-Real-Time PCR

Background: A simple and reliable DNA extraction & quantification method of hepatitis B virus (HBV) is critical in developing a viral load measurement for HBV infection. Current commercially available plasma Hepatitis B Virus (HBV), DNA extraction & quantification methods are less sensitive and require optimization, which restrict wide adoption in clinical laboratories. This study offers a report on the quantitative viral loadartus® HBV RG PCR assay by combining QIAamp® DSP Plasma DNA extraction with Rotor gene Q-Real-time PCR (Rotor gene Q-PCR). Methods: Plasma was separated by centrifugation after collection of whole peripheral blood, which was then stored at -20 C until process. DNA was extracted as per instruction & guidelines of manufacturer. The recovered eluted DNA was mixed into PCR master-mix. The Q-PCR assay performance, including linearity, diagnostic accuracy and reliability were determined. Viral load compared of the no viral load, low viral titer & high viral titer from the clinical samples for evaluation. Results: The average % yield for HBV DNA standards used was within in the range (SD 0.04 to 0.13, CV% 0.97 to 2.22). The coefficient of variation (CV) of HBV quantification for low and high titer samples was 2.23% and 2.37%, respectively. The real-time assay linearity was demonstrated with a slope of -3.03 to -3.50 and R values of 0.98 to 0.99. Accuracy and reliability of the Rotor gene Q-PCR assay was confirmed with the reference panel, and there was a strong correlation between the two results (R2 = 0.99, < 0.01) & found no major difference in observed viral load. Conclusions: The Rotorgene Q-PCR performance with artus-DSP assay is reliable and wide linear range quantification method and can be used for sensitive detection of plasma HBV with low nucleic acids content.