Amini-Bavil-Olyaee Samad, Hosseini Sayed Younes, Sabahi Farzaneh, Alavian Seyed-Moayed
Biotechnology Department, Pasteur Institute of Iran, 13164 Tehran, Iran.
Int J Infect Dis. 2008 Jan;12(1):83-7. doi: 10.1016/j.ijid.2007.05.001. Epub 2007 Aug 14.
A few reports exist on hepatitis B virus (HBV) genotype distribution in Iran; however the sample sizes of these studies are insufficient. The first objective of this study was to determine the HBV genotype distribution with a large sample size (147 specimens). The second objective was to determine the incidence of the lamivudine-resistant YMDD mutant profile among HBV-infected patients not treated with lamivudine; some studies have reported that YMDD mutants are detectable even before antiviral treatment.
We used two cost-effective PCR-based methods that have been developed in-house: gap-PCR and artificially created restriction site-PCR (ACRS-PCR). Also, 11 samples were randomly selected and bi-directionally sequenced and subjected to phylogenetic analysis.
Gap-PCR results revealed genotype D of HBV in all patients, and ACRS-PCR results disclosed the absence of mutation within the YMDD motif before antiviral therapy in the study population. Phylogenetic analysis supported the former genotyping results with the segregation of all Iranian HBV isolates in the genotype D branch with a high bootstrap value (99%, 1000 replicates).
The present study using two cost-effective methods showed that genotype D of HBV is dominant among Iranian HBV-infected subjects, and HBV lamivudine-resistant strains do not exist naturally among Iranian patients not treated with lamivudine.
关于伊朗乙型肝炎病毒(HBV)基因型分布已有一些报道;然而这些研究的样本量不足。本研究的首要目的是通过大样本量(147份标本)确定HBV基因型分布。第二个目的是确定未接受拉米夫定治疗的HBV感染患者中拉米夫定耐药YMDD突变型的发生率;一些研究报道即使在抗病毒治疗前也可检测到YMDD突变体。
我们使用了两种自行开发的具有成本效益的基于PCR的方法:缺口PCR和人工创建限制性酶切位点PCR(ACRS-PCR)。此外,随机选择11份样本进行双向测序并进行系统发育分析。
缺口PCR结果显示所有患者的HBV均为D基因型,ACRS-PCR结果表明研究人群在抗病毒治疗前YMDD基序内无突变。系统发育分析支持了先前的基因分型结果,所有伊朗HBV分离株在D基因型分支中分离,自展值较高(99%,1000次重复)。
本研究使用两种具有成本效益的方法表明,HBV D基因型在伊朗HBV感染受试者中占主导地位,且在未接受拉米夫定治疗的伊朗患者中天然不存在对拉米夫定耐药的HBV毒株。