Length variability of telomeric repeat sequences of human herpesvirus 6 DNA
Length variability of telomeric repeat sequences of human herpesvirus 6 DNA
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DOI：
10.1016/j.jviromet.2009.03.002
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发表时间：
2009-07-01
影响因子：
3.1
通讯作者：
Agut, Henri
中科院分区：
文献类型：
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作者：
Achour, Abla;Malet, Isabelle;Agut, Henri
端粒重复序列（TRS）是人类疱疹病毒6型（HHV-6）基因组中功能未知的独特结构。本研究的目的是调查TRS拷贝数在不同实验室毒株和HHV-6感染的临床标本中关于HHV-6的两种变体A和B的变异性。将从感染细胞获得的DNA进行PCR测定，该PCR测定设计用于扩增含有HHV-6A或HHV-6 B特异性TRS的基因组部分。扩增子通过琼脂糖凝胶电泳、溴化乙锭染色和核苷酸测序进行分析。TRS拷贝数在不同的实验室菌株和研究的临床标本之间差异很大，范围从15到180多。然而，对于在细胞培养物中以及在来自同一受试者的不同样品中连续繁殖后的给定菌株，该数量是恒定的。这允许在同一患者中检测到两种不同的HHV-6A菌株的混合感染。基于PCR的HHV-6 TRS分析具有有限的灵敏度，但具有高度特异性，这为将其纳入专用于HHV-6流行病学研究的分子工具集提供了机会。(C)2009爱思唯尔有限公司版权所有。
The telomeric repeat sequences (TRS) located near both ends of human herpesvirus 6 (HHV-6) genome are unique structures of unknown function among human herpesviruses. The goal of the present study was to investigate the variability of TRS copy number among different laboratory strains and HHV-6-infected clinical specimens regarding the two variants A and B of HHV-6. DNA obtained from infected cells was submitted to a PCR assay designed to amplify the part of genome containing TRS specifically either for HHV-6A or HHV-6B. Amplicons were analyzed by electrophoresis on agarose gel with ethidium bromide staining and nucleotide sequencing. The number of TRS copies was highly variable among the distinct laboratory strains and clinical specimens studied, ranging from 15 up to more than 180. However, this number was constant for a given strain after serial propagation in cell cultures as well as in different samples from the same subject. This permitted to detect a mixed infection with two distinct strains of HHV-6A within the same patient. The PCR-based analysis of HHV-6 TRS has a limited sensitivity but is highly specific, which provides the opportunity to include it in the set of molecular tools dedicated to the study of HHV-6 epidemiology. (C) 2009 Elsevier B.V. All rights reserved.