Comparison of droplet digital PCR to real-time PCR for quantification of hepatitis B virus DNA. Issue 11 (1st November 2016)

• Record Type: Journal Article
• Authors: Tang, Hui; Cai, Qingchun; Li, Hu; Hu, Peng
• Is Part Of: Bioscience, biotechnology, and biochemistry. Volume 80:Issue 11(2016)
• Page Start: 2159
• Page End: 2164
• Publication Date: 2016-11-01
• Subjects: Droplet digial PCR -- ddPCR -- real-time PCR -- HBV DNA; Biotechnology -- Periodicals; Biochemistry -- Periodicals; 660.6
• DOI: 10.1080/09168451.2016.1196576
• Languages: English
• ISSNs: 0916-8451

Abstract

Abstract: Quantitative real-time PCR (qPCR) has been widely implemented for clinical hepatitis B viral load testing, but a lack of standardization and relatively poor precision hinder its usefulness. Droplet digital PCR (ddPCR) is a promising tool that offers high precision and direct quantification. In this study, we compared the ddPCR QX100 platform by Bio-Rad with the CFX384 Touch Real-Time PCR Detection System (Bio-Rad, USA) to detect serial plasmid DNA dilutions of known concentrations as well as HBV DNA extracted from patient serum samples. Both methods showed a high degree of linearity and quantitative correlation. However, ddPCR assays generated more reproducible results and detected lower copy numbers than qPCR assays. Patient sample quantifications by ddPCR and qPCR were highly agreeable based on the Bland–Altman analysis. Collectively, our findings demonstrate that ddPCR offers improved analytical sensitivity and specificity for HBV measurements and is suitable for clinical HBV detection. Graphical abstract: : The ddPCR assay exhibited superior repeatable quantitative results over the qPCR assay, especially in settings with low copy number samples.