scholarly journals Development of a novel DNA sequencing method not only for hepatitis B virus genotyping but also for drug resistant mutation detection

2013 ◽  
Vol 6 (S1) ◽  
Author(s):  
Fanjun Wang ◽  
Lili Lu ◽  
Changshun Yu ◽  
Zhanwu Lv ◽  
Xuelian Luo ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Dna Sequencing ◽  
Mutation Detection ◽  
Drug Resistant ◽  
Sequencing Method ◽  
B Virus ◽  
Drug Resistant Mutation ◽  
Resistant Mutation

scholarly journals Hepatitis B virus pre-existing drug resistant mutation is related to the genotype and disease progression

2017 ◽  
Vol 11 (09) ◽  
pp. 727-732 ◽  
Author(s):  
Liping Wang ◽  
Fangzheng Han ◽  
Hualing Duan ◽  
Fang Ji ◽  
Xuebing Yan ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Hbv Infection ◽  
Drug Resistant ◽  
Resistance Mutations ◽  
Hbv Genotype ◽  
B Virus ◽  
Drug Resistant Mutation ◽  
Genotype C ◽  
Resistant Mutation

Introduction: Previous studies have indicated that the drug-resistant mutations of hepatitis B virus (HBV) are a major obstacle to antiviral therapy. However, it is still unclear whether there are pre-existent resistance mutations in patients with HBV infection and the relationship between drug-resistant mutation, genotypes, and progression of hepatitis B disease. Methodology: A total of 357 treatment-naïve patients with HBV infection were involved in this retrospective study. The drug-resistant mutations of HBV reverse transcriptase domain were screened by direct gene sequencing. Results: Lamivudine (LAM) resistance was detected in 8 patients (3.7%) with chronic hepatitis B (CHB), 13 (11.7%) patients with liver cirrhosis (LC), and 6 (21.4%) patients with hepatocellular carcinoma (HCC). Adefovir(ADV)-resistant mutations were detected in 10 (4.6%) patients with CHB, 15 (13.5%) patients with  LC and 4 (14.5%) patients with HCC. Both LAM and ADV resistant mutations were detected in 2 patients (0.9%) with CHB, 1 patient (0.9%) with LC and 1 patient (3.6%) with HCC. Significant differences (p <0.01) were observed in the drug-resistance rates among patients with CHB, LC and HCC. Meanwhile, all the drug-resistant mutations were found in patients with HBV genotype C. Conclusions: This study demonstrated higher risk of pre-existing drug-resistant mutations in patients with HBV genotype C comparing to patients with HBV genotype B. Likewise, increasing prevalence of pre-existing drug-resistant mutations was shown, alongside with the progression of the disease.

scholarly journals Drug-resistant mutation andgenotype of hepatitis B virus Heilongjiang Province

2005 ◽  
Vol 13 (14) ◽  
pp. 1696
Author(s):  
Di Li ◽  
Shu-Yun Zhang ◽  
Hong-Xi Gu ◽  
Xian-Feng Cheng ◽  
Xiao-Yan Wang
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Heilongjiang Province ◽  
Drug Resistant ◽  
B Virus ◽  
Drug Resistant Mutation ◽  
Resistant Mutation

scholarly journals A Deep-Sequencing Method Detects Drug-Resistant Mutations in the Hepatitis B Virus in Indonesians

Intervirology ◽  
2014 ◽  
Vol 57 (6) ◽  
pp. 384-392 ◽  
Author(s):  
Dewiyani Indah Widasari ◽  
Yoshihiko Yano ◽  
Didik Setyo Heriyanto ◽  
Takako Utsumi ◽  
Laura Navika Yamani ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Deep Sequencing ◽  
Drug Resistant ◽  
Sequencing Method ◽  
B Virus

scholarly journals Mutation Detection in the Drug-Resistant Hepatitis B Virus Polymerase Gene Using Nanostructured Reverse Micelles

2004 ◽  
Vol 20 (11) ◽  
pp. 1609-1611 ◽  
Author(s):  
Lian-Chun PARK ◽  
Tatsuo MARUYAMA ◽  
Noriho KAMIYA ◽  
Masahiro GOTO ◽  
Hiroyuki KUMA ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Reverse Micelles ◽  
Mutation Detection ◽  
Drug Resistant ◽  
Polymerase Gene ◽  
B Virus

scholarly journals Sensitivity of drug-resistant mutants of hepatitis B virus to poly-IC

2014 ◽  
Vol 58 (04) ◽  
pp. 348-355
Author(s):  
Q. ZHOU ◽  
E. CHEN ◽  
L. CHEN ◽  
Y. NONG ◽  
X. CHENG ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Drug Resistant ◽  
B Virus ◽  
Resistant Mutants

Acute hepatitis B infection associated with drug-resistant hepatitis B virus

2010 ◽  
Vol 48 (4) ◽  
pp. 270-274 ◽  
Author(s):  
Zhihui Xu ◽  
Yan Liu ◽  
Teng Xu ◽  
Li Chen ◽  
Lanlan Si ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Acute Hepatitis ◽  
Drug Resistant ◽  
Hepatitis B Infection ◽  
B Virus ◽  
Acute Hepatitis B

scholarly journals Use of Massively Parallel Ultradeep Pyrosequencing To Characterize the Genetic Diversity of Hepatitis B Virus in Drug-Resistant and Drug-Naive Patients and To Detect Minor Variants in Reverse Transcriptase and Hepatitis B S Antigen

2008 ◽  
Vol 83 (4) ◽  
pp. 1718-1726 ◽  
Author(s):  
Mariacarmela Solmone ◽  
Donatella Vincenti ◽  
Mattia Carlo Felice Prosperi ◽  
Alessandro Bruselles ◽  
Giuseppe Ippolito ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Reverse Transcriptase ◽  
Direct Sequencing ◽  
Relative Sensitivity ◽  
Massively Parallel ◽  
Drug Resistant ◽  
Resistance Mutations ◽  
B Virus ◽  
Drug Naïve

ABSTRACT Direct population sequencing and reverse hybridization (line probe assay [LiPA])-based methods are the most common methods for detecting hepatitis B virus (HBV) drug resistance mutations, although only mutations present in viral quasispecies with a prevalence of ≥20% can be detected by sequencing, and only known mutations are detected by LiPA. Massively parallel ultradeep pyrosequencing (UDPS; GS FLX platform) was used to analyze HBV quasispecies in reverse transcriptase (RT) and hepatitis B S antigen (HBsAg) from five drug-naive patients and eight drug-resistant patients. Eight primer pairs were used to obtain partially overlapping amplicons, covering the RT gene from codons 1 to 288 and the complete overlapping HBsAg sequence. A 1% mutation frequency was selected as the cutoff based on an error rate estimated on plasmid DNA. This technology enabled simultaneous analysis of between 2,852 and 18,016 clonally amplified fragments from each patient. The results indicate that UDPS has a relative sensitivity much higher than both direct sequencing and LiPA. In addition, the UDPS results are quantitative, allowing establishment of the relative frequency of both known mutations and novel substitutions. Some of the detected RT substitutions led to changes also in HBsAg. On the whole, genotype D presented a higher heterogeneity than genotype A. Considering the high quantity of information that can be provided by a single test from one patient, the short turnaround time, the information on substitution frequency, and the detection of rare variants, there are strong advantages conferred by UDPS, and the new method could play a relevant role in the clinical management of HBV infection and therapy.

In-Situ Mutation Detection by Magnetic Beads-Probe Based on Single Base Extension and Its Application in Genotyping of Hepatitis B Virus Pre-C Region 1896nt Locus Single Nucleotide Polymorphisms

2019 ◽  
Vol 15 (12) ◽  
pp. 2393-2400
Author(s):  
Xianbo Mou ◽  
Danni Sheng ◽  
Zhu Chen ◽  
Mei Liu ◽  
Yuan Liu ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Fluorescence Intensity ◽  
Magnetic Beads ◽  
Mutation Detection ◽  
Single Base Extension ◽  
Single Base ◽  
Base Extension ◽  
B Virus

Hepatitis B virus (HBV) is closely related to occurrence and development of viral hepatitis. A mutation of 1896nt locus in its pre-C region can promote replication of HBV DNA and improve stability of pre-genome RNA structure, and can even help HBV evade immune clearance. In this study, magnetic beads-probe (MBs@probe) method, combined with single base extension (SBE) technology, was developed for in-situ mutation detection of HBV pre-C region 1896nt locus. Before successfully completing the genotyping of 165 HBV samples, the crucial reaction conditions were first optimized, such as SBE temperature, MBs size and amount, and probe concentration on the surface of MBs. Experimental results showed that these conditions had significant effects on MBs@probe in-situ mutation detection. Comprehensive considerations, such as 58 °C of SBE temperature, high fluorescence intensity and signal-to-noise ratios (SNRs) were obtained when MBs@probe complex was made by 100 μg of 300 nm-MBs and 3.0 μM of probes in the system. Finally, 1896nt locus mutation in pre-C region of 165 HBV samples was successfully genotyped, among which 71 HBV samples were wild types and the remaining 94 samples were mutant types. Meanwhile, 14 randomly chosen samples were taken to further analyze fluorescence intensity and SNRs respectively, and sequencing results for the first two samples were consistent with results from the MBs@probe in-situ mutation detection method. Compared with two-color fluorescence hybridization (TCFH) genotyping technology, this method generally improves the SNRs to more than 10 (which is more than 2-fold), has higher reliability and is more suitable to detect SNPs for known sites.

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