Comparison of droplet digital PCR to real-time PCR for quantification of hepatitis B virus DNA.
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.
a Key Laboratory of Molecular Biology for Infectious Diseases (Ministry of Education), Department of Infectious Diseases , Institute for Viral Hepatitis, The Second Affiliated Hospital, Chongqing Medical University , Chongqing , P.R. China.
Hui
H
Tang
a Key Laboratory of Molecular Biology for Infectious Diseases (Ministry of Education), Department of Infectious Diseases , Institute for Viral Hepatitis, The Second Affiliated Hospital, Chongqing Medical University , Chongqing , P.R. China.
Qingchun
Q
Cai
a Key Laboratory of Molecular Biology for Infectious Diseases (Ministry of Education), Department of Infectious Diseases , Institute for Viral Hepatitis, The Second Affiliated Hospital, Chongqing Medical University , Chongqing , P.R. China.
Hu
H
Li
a Key Laboratory of Molecular Biology for Infectious Diseases (Ministry of Education), Department of Infectious Diseases , Institute for Viral Hepatitis, The Second Affiliated Hospital, Chongqing Medical University , Chongqing , P.R. China.
Peng
P
Hu
Droplet digial PCR; HBV DNA; ddPCR; real-time PCR