Hepatitis B is one of the most common infectious diseases in the world, and 350 million people have been estimated to be chronic hepatitis B virus carriers world-wide. Hepatitis B virus (HBV) has been classified into 8 genotypes (A-H) based on an intergroup divergence of 8% or more in the complete nucleotide sequence. Different genotypes of the hepatitis B virus may influence the clinical outcome of the disease. HBV genotyping method using restriction fragment length polymorphism (RFLP) can reliably identify genotypes. HBV genotyping with S gene sequence is consistent with genetic analysis using the full genomic sequences. The aim of this study was to determine the genotypes of HBV by using restriction fragment length polymorphism (RFLP) method in the region of Elazig. A total of 127 HBV-DNA positive patients (74 male, 53 female) were included in the study. Semi-nested polymerase chain reaction (PCR) was performed to amplify the specific parts of HBV S gene. In the first step, 685 base paired (bp) region was amplified by sense primer HBMF1 and anti-sense primer HBMR2, while in the second step 485 bp region was amplified by using inner-sense primer HBMF2 and anti-sense primer HBMR2. PCR products were then digested by the restriction enzymes, Alwl, Earl, Hphl, Ncil and NlalV. The RFLP assay indicated that genotype D was the only detected type in our samples. In conclusion, genotype D is the predominant type among hepatitis B patients in our region. RFLP is considered to be an easy and useful method for genotyping HBV strains.

译文

:乙型肝炎是世界上最常见的传染病之一,据估计全世界有3.5亿人是慢性乙型肝炎病毒携带者。乙型肝炎病毒(HBV)已根据完整核苷酸序列中8%或更高的组间差异分为8个基因型(A-H)。乙型肝炎病毒的不同基因型可能会影响该疾病的临床结果。使用限制性片段长度多态性(RFLP)的HBV基因分型方法可以可靠地鉴定基因型。具有S基因序列的HBV基因分型与使用完整基因组序列进行的遗传分析一致。这项研究的目的是通过使用限制性片段长度多态性(RFLP)方法在Elazig地区确定HBV的基因型。研究共纳入127例HBV-DNA阳性患者(男74例,女53例)。进行了半巢式聚合酶链反应(PCR),以扩增HBV S基因的特定部分。第一步,通过有义引物HBMF1和反义引物HBMR2扩增685个碱基对(bp)区域,而在第二步中,通过使用内感引物HBMF2和反义引物HBMR2扩增485 bp对区域。然后用限制性酶Alwl,Ear1,Hph1,Ncil和NalV消化消化PCR产物。 RFLP分析表明,基因型D是我们样本中唯一检测到的类型。总之,在我们地区的乙型肝炎患者中,基因型D是主要类型。 RFLP被认为是对HBV毒株进行基因分型的简便方法。

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