The drug treatment not only reduces HBV loads to a low level, but also results in resistance mutations in the patients. Low levels of serum HBV DNA occur in patients during drug treatment, which is considered to be one of the indicators of drug resistance in the HBV patients. The EASL guideline recommends treatment of patients with any detectable level of serum HBV DNA. The AASLD and APASL guidelines recommend antiviral therapy for patients with compensated cirrhosis and serum HBV DNA level >2000 IU/mL regardless of ALT level. The clinical efficacy of the treatment depends on various factors, such as severity of liver disease, serum ALT levels, and serum HBV DNA levels. Currently, there are 2 types of anti-HBV drugs: interferon-alpha (IFN-α) and nucleoside analogs (NAs), such as lamivudine (LAM), telbivudine (LdT), and entecavir (ETV). It has been shown to be effective in suppressing HBV replication, decreasing inflammation and fibrosis in the liver, and preventing progression of liver disease.
Antiviral therapy is an efficient way to prevent bad clinical outcomes of HBV infection.
Many drugs have been approved for the treatment of chronic hepatitis B. The estimated worldwide mortality caused by HBV infection is about 780,000 deaths a year. The patients with decompensated cirrhosis have a poor prognosis with a 14% to 35% probability of survival for 5 years. People with hepatitis B are at an increased risk of developing hepatic decompensation, cirrhosis, and hepatocellular carcinoma (HCC). There are >350 million chronic HBV carriers, 75% of whom reside in the Asia-Pacific region, especially in China. Hepatitis B virus (HBV) is one of the most serious and prevalent health problems affecting >2.4 billion people worldwide. This method is a sensitive and reliable direct sequencing method for HBV genotyping and antiviral drug resistance mutation detection, and is helpful for efficiently monitoring the response to therapy in HBV patients. We examined the reliability of the method in clinical samples, and found it could detect the HBV subtypes and drug resistance mutations in 80 clinical HBV samples with low HBV DNA levels ranging from 20 to 200 IU/mL. 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.
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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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