These primers could efficiently amplify the RT region of HBV virus at low DNA levels by directly sequencing the resulting PCR products, and mapping with the reference sequence made it possible to clearly obtain the HBV subtypes and identify the resistance mutations in the samples with HBV DNA level as low as 20 IU/mL. The novel, common, and universal primers were identified by alignment of RT region of all the HBV DNA sequences in databases. Also, sensitive detection of HBV antiviral drug resistance mutations is essential for monitoring therapy response.Īsensitive direct sequencing method for genotyping and the drug resistance mutation detection of low levels of HBV DNA in patients’ plasma is developed by PCR amplification of the DNA with novel universal primers. HBV (hepatitis B virus) genotyping is important in determining the clinical manifestation of disease and treatment response, particularly, in patients with low viral loads.
SEQUENCHER 5.2 LICENSE
This is an open-access article distributed under the terms of the Creative Commons Attribution-Non Commercial License 4.0 (CCBY-NC), where it is permissible to download, share, remix, transform, and buildup the work provided it is properly cited. This work was financially supported by the Hubei Provincial Natural Science Foundation of China (2010CDB06903), National Natural Science Foundation of China (81000771), National Key Basic Research Program of China-973 Program (2012CB526706), and the National Natural Science Foundation of China (81271694).Īll the authors in the manuscript have no conflict of interests to declare. YT, BL, and HL contributed equally to this work. BClinical Molecular Diagnostic Center, Renmin Hospital of Wuhan UniversityĬDepartment of Pathology Affiliated Tianyou Hospital of Wuhan University of Science and Technology, WuhanĭEastern Hepatobiliary Surgery Hospital, Second Military Medical University, Shanghai, ChinaĮPennsylvania State University College of Medicine and Hershey Medical center, Hershey, PAįNew York University College of Arts and Science, New York, NY.Ĭorrespondence: Yan Li, Department of Clinical Laboratory, Renmin Hospital of Wuhan University, 99 Ziyang Road of Wuchang District, Wuhan 430060, China (e-mail: ) Chunhua Song, Pennsylvania State University College of Medicine, Penn State Hershey Children's Hospital, PO Box 850, 500 University Drive, Hershey, PA 17033 (e-mail: ).Ībbreviations: ETV = entecavir, HBV = hepatitis B virus, HCC = hepatocellular carcinoma, IFN-a = interferon-alpha, LAM = lamivudine, LdT = telbivudine, NAs = nucleoside analogs, qPCR = quantitative real-time PCR.
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