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基于乙肝病毒DNA聚合酶基因多态性对印度西部慢性乙肝患者基因型的预测

Hepatitis B virus DNA polymerase gene polymorphism based prediction of genotypes in chronic HBV patients from Western India.

作者信息

Chavan Yashwant G, Pawar Sharad R, Wani Minal, Raut Amol D, Misra Rabindra N

机构信息

Dr. D Y Patil Biotechnology and Bioinformatics Institute, Tathwade, Pune 411033. Dr. D Y Patil Vidyapeeth, Sant Tukaram Nagar, Pimpri, Pune 411018. Maharashtra, India.

Medical Genetics, geneOmbio Technologies Private Limited, Baner, Pune 411045. Maharashtra, India.

出版信息

Afr Health Sci. 2017 Sep;17(3):762-772. doi: 10.4314/ahs.v17i3.19.

Abstract

BACKGROUND

Hepatitis B Virus (HBV) infection is one of the major causes of liver cirrhosis, hepatocellular carcinoma and deaths due to the acute or chronic consequences worldwide. HBV is distributed into various genotypes based on nucleic acid sequence variation.

OBJECTIVES

To develop a method of HBV genotyping and drug resistance interpretation using partial sequencing of polymerase gene.

METHODS

This study was performed on 98 HBV infected patients' serum samples from Western India. A nested PCR protocol was designed for amplification of pol gene from HBV genome and Sanger's sequencing of the gene fragment. Sequences were aligned with HBV reference sequences for phylogenetic analysis and for characterization of genetic diversity. Drug resistance mutations were screened using HBVSeq program from Stanford University.

RESULTS

Distribution of HBV genotypes showed predominance of genotype D, circulating in 76 (77.55%) patients (p < 0.05). Genotypes A and C were less prevalent and were identified in 4 (4.08%) and 18 (18.37%) patients, respectively. Anti-retroviral drug resistance mutations were not detected in any patient.

CONCLUSION

A method for determination of HBV genotypes using pol gene sequencing which simultaneously detects major drug resistance mutations has been established. HBV genetic diversity may play an important role in treatment decision.

摘要

背景

乙肝病毒(HBV)感染是全球范围内导致肝硬化、肝细胞癌以及因急慢性后果所致死亡的主要原因之一。基于核酸序列变异,HBV可分为多种基因型。

目的

利用聚合酶基因的部分测序开发一种HBV基因分型及耐药性解读方法。

方法

本研究对来自印度西部的98例HBV感染患者的血清样本进行检测。设计了一种巢式PCR方案,用于从HBV基因组中扩增pol基因,并对基因片段进行桑格测序。将序列与HBV参考序列进行比对,以进行系统发育分析和遗传多样性特征分析。使用斯坦福大学的HBVSeq程序筛选耐药突变。

结果

HBV基因型分布显示D基因型占主导,76例(77.55%)患者中存在该基因型(p < 0.05)。A基因型和C基因型较少见,分别在4例(4.08%)和18例(18.37%)患者中检出。未在任何患者中检测到抗逆转录病毒药物耐药突变。

结论

已建立一种利用pol基因测序测定HBV基因型的方法,该方法可同时检测主要耐药突变。HBV遗传多样性可能在治疗决策中起重要作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1ca5/5656201/dabd505b0579/AFHS1703-0762Fig1.jpg

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