Comparison of droplet digital PCR to real-time PCR for quantification of hepatitis B virus DNA
Comparison of droplet digital PCR to real-time PCR for quantification of hepatitis B virus DNA
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DOI：
10.1080/09168451.2016.1196576
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发表时间：
2016-01-01
影响因子：
1.6
通讯作者：
Hu, Peng
中科院分区：
文献类型：
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作者：
Tang, Hui;Cai, Qingchun;Hu, Peng
实时定量PCR（qPCR）已广泛应用于临床B型肝炎病毒载量检测，但缺乏标准化和相对较差的精确度阻碍了其实用性。液滴数字PCR（ddPCR）是一种有前途的工具，提供高精度和直接定量。在这项研究中，我们比较了Bio-Rad的ddPCR QX 100平台与CFX 384 Touch实时PCR检测系统（Bio-Rad，USA），以检测已知浓度的系列质粒DNA稀释液以及从患者血清样本中提取的HBV DNA。两种方法均表现出高度的线性和定量相关性。然而，ddPCR测定产生的结果更具重现性，并且检测到的拷贝数低于qPCR测定。基于Bland-Altman分析，通过ddPCR和qPCR进行的患者样本定量高度一致。总的来说，我们的研究结果表明，ddPCR提供了改善的分析灵敏度和特异性的HBV测量，适用于临床HBV检测。
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.