New suggestions for optimization, validation of the quantitative measurement of HBV cccDNA

Approximately 300 million individuals worldwide are chronically contaminated with the hepatitis B virus (HBV) and at excessive danger of growing liver cirrhosis or hepatocellular carcinoma. Consequently, there’s an pressing want for the event of HBV healing therapies. Due to the distinctive replication technique of the HB virus, nevertheless, quantification of viral DNA in contaminated liver cells, which is crucial in preclinical and scientific research, is technically troublesome and never standardised. An worldwide analysis consortium led by DZIF scientists has now developed suggestions for the optimisation, management and validation of quantitative viral DNA measurements, which may very well be essential for preclinical and scientific analysis of therapeutic methods.

HBV is a DNA virus that particularly infects human hepatocytes. After an infection, the DNA genome of the virus is transformed by mobile enzymes into covalently closed round DNA (cccDNA), which types secure mini chromosomes throughout the cell nuclei by affiliation with mobile chromosomal proteins. In this kind, the viral DNA then serves as a template for the formation of latest virus particles. A significant limitation of preclinical and scientific HBV analysis is the shortage of standardised PCR-based strategies for the precise quantification of viral DNA current as cccDNA in HBV-infected samples. As a part of a world consortium supported by the International Coalition to Eliminate HBV (ICE-HBV), DZIF scientists on the University Medical Center Hamburg-Eppendorf, Heidelberg University and the Technical University of Munich labored along with the French L’Agence nationale de recherches sur le sida et les hépatites virales (ANRS) and colleagues at Gilead Sciences within the USA to check protocols and develop evidence-based greatest practices steering to quantify cccDNA by quantitative PCR.

“When evaluating completely different protocols for quantifying HBV DNA in tissue samples, we discovered that the coexistence of various types of viral DNA in addition to circumstances of pattern preservation and remedy strongly have an effect on PCR-based quantification of HBV cccDNA,” says DZIF younger investigator Dr Lena Allweiss, who led the collaboration of the six taking part laboratories. Based on the research outcomes, the workforce developed pattern type-tailored methodological suggestions for the optimisation, management and validation of the quantitative measurement of HBV cccDNA.

“The research will help the HBV remedy analysis programmes aiming at assessing the affect of therapies on the mobile HBV reservoir in preclinical research and scientific trials,” sums up research lead Prof. Maura Dandri, DZIF scientist on the University Medical Center Hamburg-Eppendorf, the outcomes of the research.