title: A novel real-time reverse transcription-polymerase chain reaction assay with partially double-stranded linear DNA probe for sensitive detection of hepatitis C viral RNA.
abstract: The detection and quantification of HCV RNA is very helpful for the management and treatment of HCV related diseases. Detection of low HCV viral load is a great challenge in HCV RNA detection. Here, we developed a novel real-time RT-PCR assay with partially double-stranded linear DNA probe which can detect all HCV genotypes and improve the detection performance. The novel assay has a wide linear dynamic range of HCV RNA quantification (1×10(2)-1×10(11)IU/ml) and a limit of detection of 78IU/ml. The assay exhibits an excellent reproducibility with 2.52% and 1.33% coefficients of variations, for inter- and intra-assays, respectively. To evaluate the viability of the assay, a comparison with a commercial HCV RNA detection kit was performed using 106 serum samples. The lineared correlation coefficient between the novel assay and the commercial HCV RNA detection kit was 0.940. Meanwhile, the deviation between the two methods was tolerable. Therefore, the novel real-time RT-PCR assay was applicable for laboratory diagnosis and monitoring of HCV infection.
authors.affiliation: Department of Hepatology, Lanzhou University Second Hospital, Lanzhou, Gansu, China; Pathogen Diagnostic Center, CAS Key Laboratory of Molecular Virology & Immunology, Institut Pasteur of Shanghai, Chinese Academy of Science, Shanghai, China.
authors.forename: Tianfu
authors.initials: T
authors.lastname: Liu
authors.affiliation: Medical Laboratory of Taizhou Fourth People's Hospital, Taizhou, Jiangsu, China.
authors.forename: Zhenzhou
authors.initials: Z
authors.lastname: Wan
authors.affiliation: Pathogen Diagnostic Center, CAS Key Laboratory of Molecular Virology & Immunology, Institut Pasteur of Shanghai, Chinese Academy of Science, Shanghai, China.
authors.forename: Jia
authors.initials: J
authors.lastname: Liu
authors.affiliation: Department of Hepatology, Lanzhou University Second Hospital, Lanzhou, Gansu, China.
authors.forename: Lingyi
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Shaanxi Engineering and Technological Research Center for Conversation and Utilization of Regional Biological Resources, College of Life Sciences, Yan'an University, Yan'an, Shaanxi, China.
authors.forename: Yanheng
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: Pathogen Diagnostic Center, CAS Key Laboratory of Molecular Virology & Immunology, Institut Pasteur of Shanghai, Chinese Academy of Science, Shanghai, China.
authors.forename: Ke
authors.initials: K
authors.lastname: Lan
authors.affiliation: Pathogen Diagnostic Center, CAS Key Laboratory of Molecular Virology & Immunology, Institut Pasteur of Shanghai, Chinese Academy of Science, Shanghai, China. Electronic address: yhhu@ips.ac.cn.
authors.forename: Yihong
authors.initials: Y
authors.lastname: Hu
authors.affiliation: Pathogen Diagnostic Center, CAS Key Laboratory of Molecular Virology & Immunology, Institut Pasteur of Shanghai, Chinese Academy of Science, Shanghai, China. Electronic address: zhangcy1999@ips.ac.cn.
authors.forename: Chiyu
authors.initials: C
authors.lastname: Zhang
mesh_terms: D015342:DNA Probes / Q000235:genetics*; D016174:Hepacivirus / Q000235:genetics / Q000302:isolation & purification*; D006526:Hepatitis C / Q000175:diagnosis / Q000821:virology*; D006801:Humans; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
chemical_list: D015342:DNA Probes; D012367:RNA, Viral
keywords: HCV; Partially double-stranded linear DNA probe; Real-time RT-PCR; Viral load