Altuğlu Imre, Ozacar Tijen, Sertoz Ruchan Yazan, Erensoy Selda
Ege University Medical Faculty, Department of Microbiology and Clinical Microbiology, Bornova Izmir, Turkey.
Int J Infect Dis. 2007 Jan;11(1):58-62. doi: 10.1016/j.ijid.2005.10.012. Epub 2006 May 4.
Analysis of hepatitis delta virus (HDV) isolates from around the world has indicated that there are at least three phylogenetically distinct genotypes with different geographic distributions. The aim of this study was to determine the distribution of HDV genotypes by direct sequencing in patients with chronic delta hepatitis in Izmir, Turkey.
Serum samples from 32 chronic hepatitis patients (21 males, 11 females; mean age 44.2 years, range 23-70 years) with anti-delta positivity were analyzed for hepatitis B and C serologies. After reverse transcription, cDNA of partial delta antigen was amplified by in-house nested PCR. The products of the HDV PCR were bidirectionally sequenced with internal primers using Big Dye Terminator DNA Sequencing Kit (Applied Biosystems, CA, USA) and ABI Prism 310 Genetic Analyzer (Perkin Elmer, USA). Nucleotide sequences of HDV were compared with previously reported sequences and aligned by using ClustalW (1.82).
HDV-RNA was positive in 26 (81.3%) of 32 anti-delta positive samples. Comparison of the HDV sequences with published sequences of HDV genotypes I, II, and III indicated that all were closely related to HDV genotype I isolates. Similarity among isolated sequences ranged from 84% to 96%.
HDV genotyping was successfully performed by direct sequencing of the amplicons obtained from routine HDV-RNA screening PCR tests. All of the HDV isolates from the chronic delta hepatitis patients included in this study were found to be genotype I.
对来自世界各地的丁型肝炎病毒（HDV）分离株进行分析表明，至少存在三种系统发育上不同的基因型，且具有不同的地理分布。本研究的目的是通过直接测序确定土耳其伊兹密尔慢性丁型肝炎患者中HDV基因型的分布。
对32例抗-HDV阳性的慢性肝炎患者（21例男性，11例女性；平均年龄44.2岁，范围23 - 70岁）的血清样本进行乙肝和丙肝血清学分析。逆转录后，使用内部巢式聚合酶链反应（PCR）扩增部分δ抗原的互补DNA（cDNA）。HDV PCR产物使用Big Dye Terminator DNA测序试剂盒（美国加利福尼亚州应用生物系统公司）和ABI Prism 310遗传分析仪（美国珀金埃尔默公司），用内部引物进行双向测序。将HDV的核苷酸序列与先前报道的序列进行比较，并使用ClustalW（1.82）进行比对。
32例抗-HDV阳性样本中有26例（81.3%）HDV-RNA呈阳性。将HDV序列与已发表的HDV基因型I、II和III的序列进行比较表明，所有序列均与HDV基因型I分离株密切相关。分离序列之间的相似性范围为84%至96%。
通过对常规HDV-RNA筛查PCR试验获得的扩增子进行直接测序，成功进行了HDV基因分型。本研究中纳入的慢性丁型肝炎患者的所有HDV分离株均为基因型I。
