Rendón Julio-C, Cortés-Mancera Fabián, Duque-Jaramillo Alejandra, Ospina Marta C, Navas María Cristina
Grupo de Gastrohepatología, Facultad de Medicina, Universidad de Antioquia, Medellín, Colombia.
Biomedica. 2015 Dec 7;36(0):79-88. doi: 10.7705/biomedica.v36i0.2976.
Ten viral genotypes (A-J) distributed in all continents have been described for hepatitis B virus (HBV). One of the methodologies for determining the viral genotype is the restriction fragment length polymorphism (RFLP) technique, a simple and relatively inexpensive method, albeit with some limitations.
The initial objective of the project was to identify the HBV genotypes by RFLP in serum samples obtained from patients and blood donors. However, due to the discrepancies of RFLP patterns it was also necessary to perform phylogenetic genotyping and in silico analysis of HBV sequences.
We obtained 56 serum samples. DNA extraction was followed by PCR amplification of a fragment of HBV ORF S. We analyzed PCR products by RFLP with AlwI, BsrI, CfrI, HpaII and StyI, and we sequenced some. We compared the patterns obtained with those in previous reports. We also performed RFLP analysis in silico since we found differences between the patterns expected and those obtainedResults: We identified genotypes A and F, subgenotype F3, in the samples. This result is in agreement with those of previous studies carried out in Colombia; indeed, subgenotype F3 is the most frequent in the Andean region of the country, while genotype A is the most frequent HBV genotype in the western region (department of Chocó). Based on the in silico analysis of 229 HBV sequences from GenBank and 11 sequences of this study, we identified the RLFP pattern for genotype F, subgenotype F3, and we described some modifications of genotype A RFLP patterns.
We identified the single nucleotide polymorphism pattern for genotype F, subgenotype F3, by in silico analysis and sequencing. Further robust in silico analyses are necessary to validate the RFLP patterns of HBV genotype and subgenotypes.
已描述了分布于各大洲的10种乙型肝炎病毒（HBV）基因型（A - J）。确定病毒基因型的方法之一是限制性片段长度多态性（RFLP）技术，这是一种简单且相对便宜的方法，尽管存在一些局限性。
该项目的最初目标是通过RFLP技术在从患者和献血者获得的血清样本中鉴定HBV基因型。然而，由于RFLP模式存在差异，因此还需要对HBV序列进行系统发育基因分型和计算机模拟分析。
我们获得了56份血清样本。提取DNA后，对HBV ORF S片段进行PCR扩增。我们用AlwI、BsrI、CfrI、HpaII和StyI通过RFLP分析PCR产物，并对一些产物进行测序。我们将获得的模式与先前报告中的模式进行比较。由于我们发现预期模式与获得的模式之间存在差异，因此我们还进行了计算机模拟RFLP分析。
我们在样本中鉴定出基因型A和F以及F3亚型。这一结果与之前在哥伦比亚进行的研究结果一致；事实上，F3亚型是该国安第斯地区最常见的亚型，而基因型A是西部地区（乔科省）最常见的HBV基因型。基于对来自GenBank的229条HBV序列和本研究的11条序列的计算机模拟分析，我们确定了基因型F、F3亚型的RLFP模式，并描述了基因型A的RFLP模式的一些修饰。
我们通过计算机模拟分析和测序确定了基因型F、F3亚型的单核苷酸多态性模式。需要进一步进行有力的计算机模拟分析以验证HBV基因型和亚型的RFLP模式。
