Development of Magnetic Capture Hybridization and Quantitative Polymerase Chain Reaction for Hepatitis B Virus Covalently Closed Circular DNA
| Author | Yongcan Guo | en |
| Author | Shangchun Sheng | en |
| Author | Bin Nie | en |
| Author | Zhiguang Tu | en |
| Issued Date | 2015-01-30 | en |
| Abstract | Background: Hepatitis B virus (HBV) covalently closed circular DNA (cccDNA) served as a vital role in the life cycle of the virus and persistent infection. However, specific and quantitative methods for cccDNA detection have not been available. Objectives: Our aim was to develop and primarily evaluate a quantitative method for HBV cccDNA based on magnetic capture hybridization and quantitative PCR technology. Materials and Methods: The functionalized-nanoparticles specifically to capture HBV cccDNA, located on both sides of relaxed circle DNA (rcDNA) gap, were designed. Then, magnetic capture hybridization and quantitative PCR (MCH-qPCR) assay were developed and its performance was primarily evaluated with cccDNA standards and serum samples of patients with chronic hepatitis B. Results: Specific nanoparticles of cccDNA capture were prepared and a magnetic capture hybridization and quantitative assay method for cccDNA was developed successfully. The limit of detection was 90 IU/mL, and a good linear relationship in the range of 102-106 IU/mL was revealed (r2 = 0.994) with the MCH-qPCR. Compared with directly real-time PCR, a high content of HBV DNA did not affect the detection of cccDNA for the MCH-qPCR method, and there was no cross-reactivity between cccDNA and rcDNA. Conclusions: The novel MCH-qPCR method has good sensitivity and specificity. It could meet the requirement of clinical routine detection. | en |
| DOI | https://doi.org/10.5812/hepatmon.23729 | en |
| Keyword | Hepatitis B Virus | en |
| Keyword | Magnetic | en |
| Keyword | PCR Technology | en |
| Publisher | Brieflands | en |
| Title | Development of Magnetic Capture Hybridization and Quantitative Polymerase Chain Reaction for Hepatitis B Virus Covalently Closed Circular DNA | en |
| Type | Research Article | en |
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