Studying the genetic diversity of the varicella-zoster virus in selected regions of the Russian Federation using high-throughput sequencing

Maksim I. Nadtoka, Vladislav G. Lysenkov, M. R. Agletdinov, Andrey A. Mishkin, N. M. Afonina, A. Ploskireva, Irina V. Mikheeva, Kamil F. Khafizov, V. Akimkin
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Abstract

Introduction. Varicella-zoster virus (VZV), the causative agent of the disease of the same name and herpes zoster, is phylogenetically divided into 8 clades, the distribution of which is characterized by geographic reference to certain regions of the world. For most countries, VZV clades circulating in their territories have been identified, however, such information is almost unavailable for Russia. The purpose of the study is to develop an effective method for VZV typing using high-throughput sequencing technologies to identify the prevalence of various VZV clades in Moscow, Moscow Region, and Stavropol Territory. Materials and methods. To genotype VZV, it is enough to refer to 7 nucleotide positions. Their unique combinations can be used to assign the virus to one of the clades. Short sections of nucleotide sequences of open reading frames were obtained using a developed set of primers. Results. A VZV genotyping technique has been developed and optimized. Using this technique, primary data on the distribution of VZV clades in the studied regions have been obtained. Thus, it has been established that in Moscow and a number of other regions, the 1st, 3rd, and 5th clades of VZV are predominantly distributed. Conclusion. The developed technique, including a primer panel and a genotyping algorithm, allows VZV typing in a short time while reducing specimen preparation costs and simultaneously increasing the number of specimens in one sequencing cycle. The results obtained using this assay allow us to assume that in Moscow, Moscow Region, Stavropol Territory, VZV, clades 1, 3, and 5 are the most represented ones. To confirm this hypothesis, it is necessary to include a larger number of clinical specimens in subsequent studies, including from other regions of the country.
利用高通量测序技术研究俄罗斯联邦部分地区水痘-带状疱疹病毒的遗传多样性
导言。水痘-带状疱疹病毒(VZV)是同名疾病和带状疱疹的病原体,在系统发育上分为 8 个支系,其分布特点是以世界某些地区为地理参照。大多数国家已经确定了在其境内流行的 VZV 支系,但俄罗斯几乎没有此类信息。 本研究的目的是利用高通量测序技术开发一种有效的 VZV 分型方法,以确定各种 VZV 支系在莫斯科、莫斯科州和斯塔夫罗波尔边疆区的流行情况。 材料和方法。要对 VZV 进行基因分型,只需参考 7 个核苷酸位置即可。它们的独特组合可用于将病毒归入其中一个支系。使用一套开发的引物获得了开放阅读框的核苷酸序列短片。 结果开发并优化了 VZV 基因分型技术。利用这一技术,获得了研究地区 VZV 支系分布的原始数据。由此确定,在莫斯科和其他一些地区,主要分布着 VZV 的第 1、第 3 和第 5 支系。 结论所开发的技术(包括引物板和基因分型算法)可在短时间内对 VZV 进行分型,同时降低标本制备成本,并在一个测序周期内增加标本数量。利用这种检测方法获得的结果使我们可以推测,在莫斯科、莫斯科州、斯塔夫罗波尔边疆区,VZV 1、3 和 5 支系是最具代表性的支系。为了证实这一假设,有必要在后续研究中纳入更多的临床标本,包括来自国内其他地区的标本。
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