Clinical implications of mutations C-to-T1653 and T-to-C/A/G1753 of hepatitis B virus genotype C genome in chronic liver disease

Takahashi, Kazuaki; Ohta, Yasuhiko; Kanai, Koichi; Akahane, Yoshihiro; Iwasa, Yasushi; Hino, Keisuke; Ohno, Naofumi; Yamada, Hiroshi; Mishiro, Shunji

doi:10.1007/s007050050588

Cited by 67 publications

(60 citation statements)

References 20 publications

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“…Unfortunately, these methods are time consuming, labor intensive, and difficult to automate because they require a postamplification procedure, such as restriction enzyme digestion and electrophoresis. 21,22 The results of the present study indicate that real-time PCR with fluorescent hybridization probes represents a promising means of accurately identifying HBV mutants in patients with chronic HBV infections. Because the amount of PCR product amplified is monitored in each cycle and visualized, the technique provides rapid analyses of absolute template amounts without post-PCR steps.…”

Section: Discussionmentioning

confidence: 74%

Rapid detection of hepatitis B virus mutations using real-time PCR and melting curve analysis

2002

Hepatology

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Current methods of detecting hepatitis B virus (HBV) mutations are time consuming, labor intensive, and not suitable for screening large numbers of samples. In the present study, we documented the advantages of a system that exploits differences in thermal stability between perfect match and mismatch hybrids, and thereby distinguishes between wild-type and mutants. Hybridization probes were designed complementary to specific wild-type HBV sequences in surface (S), precore, and basal core promoter (BCP) regions of the HBV genome (nt 587, 1896, and 1762/1764, respectively). Two probes were designed for each mutation: anchor probes were 3 labeled with fluorescein and sensor probes, 5 labeled with LC-Red 640, and 3 phosphorylated. Temperatures for each probe melted from amplification products were then determined in a melting program. Sera from 12 patients, each containing identified HBV mutants (6 S-escape, 1 precore, 1 BCP, and 4 mixed precore and BCP), and 5 control sera from patients with wild-type virus were analyzed. Genomic sequences of mutant and wild-type viruses were confirmed by direct sequencing. Real-time polymerase chain reaction (PCR) with fluorescent hybridization probes accurately identified each mutant and wild-type genome. Melting temperatures obtained from probe-product duplexes for the 3 mutants were distinguished from wild-type (>4.0°C, minimal) within 45 minutes. The sensitivity of the system was 100 copies/mL and as few as 5% of mutant among wild-type virus were detected. In conclusion, real-time PCR with fluorescent hybridization probes is a specific, sensitive, quantitative, and rapid means of detecting clinically relevant HBV mutants. (HEPATOLOGY 2002;36:723-728.) T he hepatitis B virus (HBV) is one of the most common infectious agents in humans and a major cause of chronic hepatitis, cirrhosis, and hepatocellular carcinoma. [1][2][3] It is a partially double-stranded DNA molecule virus that replicates through an RNA intermediate by reverse transcription. 4,5 The reverse transcriptase enzyme responsible for this aspect of the replication cycle lacks proofreading function and, therefore, mutations to the genome are common. Indeed, it is now recognized that the majority of chronic HBV infections are associated with emergence of mutations throughout the viral genome. Some of these mutations result in the generation of diverse viral populations and different clinical outcomes. Those in the surface, core, precore, and basal core promoter (BCP) regions appear to have the greatest clinical relevance. 6,7 Additional mutations have also been described after therapeutic interventions such as antiviral therapy and vaccination. 8,9 Thus, the ability to detect these mutants is essential to further our understanding of the natural history of HBV and its response to antiviral therapy and immunoprophylaxis.Presently, the detection of HBV mutants is largely performed by restriction fragment length polymorphism analysis, 10,11 gap ligase chain reaction assay and/or direct sequencing. 12,13 These methods ...

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Section: Discussionmentioning

confidence: 74%

Rapid detection of hepatitis B virus mutations using real-time PCR and melting curve analysis

2002

Hepatology

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“…In addition, analysis of the nucleotide at position 1753 showed that a T-to-V (A/G/C) mutation increased to 46.8% in LC from 18.0% in CH, but dramatically decreased in HCC (22.9%) ( Table 4), suggesting that this mutation is associated with liver cirrhosis rather than HCC. In contrast, analysis of sera or plasma from Japanese subjects with AC, CH, LC and HCC infected with HBV genotype C showed that the percentage of T1753V mutation increased with progression of liver disease [41] . It is also reported that T1753V mutation was higher in HCC (53.2%) compared with LC (18.8%) and CH (9.8%) [31] .…”

Section: Discussionmentioning

confidence: 80%

Hepatitis B virus subgenotypes and basal core promoter mutations in Indonesia

Utama¹,

Purwantomo

Siburian

et al. 2009

WJG

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AIM:To identify the distribution of hepatitis B virus (HBV) subgenotype and basal core promoter (BCP) mutations among patients with HBV-associated liver disease in Indonesia. METHODS:Patients with chronic hepatitis (CH, n = 61), liver cirrhosis (LC, n = 62), and hepatocellular carcinoma (HCC, n = 48) were included in this study.HBV subgenotype was identified based on S or preS gene sequence, and mutations in the HBx gene including the overlapping BCP region were examined by direct sequencing. RESULTS:HBV genotype B (subgenotypes B2, B3, B4, B5 and B7) the major genotype in the samples, accounted for 75.4%, 71.0% and 75.0% of CH, LC and HCC patients, respectively, while the genotype C (subgenotypes C1, C2 and C3) was detected in 24.6%, 29.0%, and 25.0% of CH, LC, and HCC patients, respectively. Subgenotypes B3 (84.9%) and C1 (82.2%) were the main subgenotype in HBV genotype B and C, respectively. Serotype adw2 (84.9%) and adrq+ (89.4%) were the most prevalent in HBV genotype B and C, respectively. Double mutation (A1762T/G1764A) in the BCP was significantly higher in LC (59.7%) and HCC (54.2%) than in CH (19.7%), suggesting that this mutation was associated with severity of liver disease. The T1753V was also higher in LC (46.8%), but lower in HCC (22.9%) and CH (18.0%), suggesting that this mutation may be an indicator of cirrhosis. CONCLUSION:HBV genotype B/B3 and C/C1 are the major genotypes in Indonesia. Mutations in BCP, such as A1762T/G1764A and T1753V, might have an association with manifestations of liver disease.

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“…In fact, T1753A/G/C mutations are usually present along with the existence of A1762T/ G1764A mutations. Previous data also demonstrated that T1753A/G/C mutations occurred later than the double BCP mutations in the natural course of chronic HBV infection (Takahashi et al, 1999). Regarding pre-S deletion, it has been shown in several reports that such mutant is an important risk factor of HCC (Chen et al, 2006;Choi et al, 2007).…”

Section: Discussionmentioning

confidence: 94%

Hepatitis B Virus Genetic Variation and TP53 R249S Mutation in Patients with Hepatocellular Carcinoma in Thailand

Thongbai

Sa-nguanmoo²,

Kranokpiruk³

et al. 2013

Asian Pacific Journal of Cancer Prevention

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Chronic hepatitis B virus (HBV) infection and dietary exposure to aflatoxin B1 (AFB1) are major risk factors for hepatocellular carcinoma (HCC). The aim of this study was to evaluate the role of HBV genetic variation and the R249S mutation of the p53 gene, a marker of AFB1-induced HCC, in Thai patients chronically infected with HBV. Sixty-five patients with and 89 patients without HCC were included. Viral mutations and R249S mutation were characterized by direct sequencing and restriction fragment length polymorphism (RFLP) in serum samples, respectively. The prevalences of T1753C/A/G and A1762T/G1764A mutations in the basal core promotor (BCP) region were significantly higher in the HCC group compared to the non-HCC group. R249S mutation was detected in 6.2% and 3.4% of the HCC and non-HCC groups, respectively, which was not significantly different. By multiple logistic regression analysis, the presence of A1762T/G1764A mutations was independently associated with the risk of HCC in Thai patients.

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Clinical implications of mutations C-to-T1653 and T-to-C/A/G1753 of hepatitis B virus genotype C genome in chronic liver disease

Cited by 67 publications

References 20 publications

Rapid detection of hepatitis B virus mutations using real-time PCR and melting curve analysis

Rapid detection of hepatitis B virus mutations using real-time PCR and melting curve analysis

Hepatitis B virus subgenotypes and basal core promoter mutations in Indonesia

Hepatitis B Virus Genetic Variation and TP53 R249S Mutation in Patients with Hepatocellular Carcinoma in Thailand

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