单位：[1]National Center for Clinical Laboratories, Beijing Hospital, Beijing[2]Graduate School, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing[3]Department of Rheumatology,China-Japan Friendship Hospital, Beijing, People’s Republic of China
Viral nucleic acids are unstable when improperly collected, handled, and stored, resulting in decreased sensitivity of currently available commercial quantitative nucleic acid testing kits. Using known unstable hepatitis C virus RNA, we developed a quantitative RT-PCR method based on a new primer design strategy to reduce the impact of nucleic acid instability on nucleic acid testing. The performance of the method was evaluated for Linearity, limit of detection, precision, specificity, and agreement with commercial hepatitis C virus assays. Its clinical application was compared to that of two commercial kits Cobas AmpliPrep/Cobas TaqMan (CAP/CTM) and Kehua. The quantitative RT-PCR method delivered a good performance, with a linearity of R-2 = 0.99, a total Limit of detection (genotypes 1 to 6) of 42.6 IU/mL (95% CI, 32.84 to 67.76 IU/mL), a CV of 1.06% to 3.34%, a specificity of 100%, and a high concordance with the CAP/CTM assay (R-2 = 0.97), with a means +/- SD value of -0.06 +/- 1.96 log IU/mL (range, -0.38 to 0.25 tog IU/mL). The method was superior to commercial assays in detecting unstable hepatitis C virus RNA (P < 0.05). This quantitative RT-PCR method can effectively eliminate the influence of RNA instability on nucleic acid testing. The principle of primer design strategy may be applied to the detection of other RNA or DNA viruses.
基金：
program 863 from the National High Technology Research and Development Program of China [2013ZX10004805]
第一作者单位：[1]National Center for Clinical Laboratories, Beijing Hospital, Beijing[2]Graduate School, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing
共同第一作者：
通讯作者：
通讯机构：[1]National Center for Clinical Laboratories, Beijing Hospital, Beijing[2]Graduate School, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing[*1]National Center for Clinical Laboratories, Beijing Hospital, 1 Dahua Rd., Dongdan, Beijing, People’s Republic of China.
推荐引用方式(GB/T 7714)：
Chen Lida,Li Wenli,Zhang Kuo,et al.Hepatitis C Virus RNA Real-Time Quantitative RT-PCR Method Based on a New Primer Design Strategy[J].JOURNAL of MOLECULAR DIAGNOSTICS.2016,18(1):84-91.doi:10.1016/j.jmoldx.2015.07.009.
APA：
Chen, Lida,Li, Wenli,Zhang, Kuo,Zhang, Rui,Lu, Tian...&Li, Jinming.(2016).Hepatitis C Virus RNA Real-Time Quantitative RT-PCR Method Based on a New Primer Design Strategy.JOURNAL of MOLECULAR DIAGNOSTICS,18,(1)
MLA：
Chen, Lida,et al."Hepatitis C Virus RNA Real-Time Quantitative RT-PCR Method Based on a New Primer Design Strategy".JOURNAL of MOLECULAR DIAGNOSTICS 18..1(2016):84-91
