Abstract
An unprecedented on its scale Ebola fever epidemic, caused by Zaire ebolavirus, has been registered in the territory of West Africa in 2014-2015. The virus quickly spread from the Republic of Guinea into the neighboring countries. Presented are the results of comparative analysis between three Zaire ebolavirus genomes, the complementary DNA of which was obtained from the samples from Ebola fever patients in the Republic of Guinea in 2014. The nucleotide sequences of the genomes received have allowed for identification of phylogenetic affinity to certain references, displayed in the database of NCBI GenBank. General phylogenetic analysis by means of representative sampling has revealed two trends of genome evolutionary development from one and the same ancestral form. The first branch of evolution is represented by a relatively small number of Zaire ebolavirus isolates, disseminated in Guinea. The second one, the most numerous trend, is generated by four variants of Zaire ebolavirus genome, which have been identified starting from May - early June, 2014 in Sierra-Leone, but within a short period of time they have won the whole territory of the country and the neighboring Guinea and Liberia.
Highlights
An unprecedented on its scale Ebola fever epidemic, caused by Zaire ebolavirus, has been registered in the territory of West Africa in 2014–2015
The virus quickly spread from the Republic of Guinea into the neighboring countries
Presented are the results of comparative analysis between three Zaire ebolavirus genomes, the complementary DNA of which was obtained from the samples from Ebola fever patients in the Republic of Guinea in 2014
Summary
Получение необходимых фрагментов ДНК для секвенирования проводили в два этапа, используя гнездовую ПЦР. Секвенирование полученных специфичных фрагментов ДНК проводили с использованием генетического анализатора «ABI 3500xl» (LifeTechnologies), согласно стандартному протоколу производителя. Часть фрагментов генома визуально не детектируемых при учете результатов ПЦР в агарозном геле секвенировали после целевого обогащения исходной кДНК, используя систему высокопроизводительного секвенирования PGM (IonTorrent), согласно стандартному протоколу производителя. Фрагменты генома после секвенирования собирали в единую последовательность с помощью выравнивая на референс – изолят Makona-Kissidougou-C15 (KJ660346, 17.03.2014, Гвинея) с использованием программ SeqMan Pro 11 Для проведения сравнительного анализа полученных данных нами использована репрезентативная выборка из международной базы данных NCBI GenBank, содержащая 645 геномов ZE, полученных из клинического материала от больных лихорадкой Эбола на территории стран Западной Африки (Гвинейская Республика, Сьерра-Леоне, Либерия и Мали), взятого с марта 2014 по март 2015 года. Для построения филогенетических дендрограмм и дальнейшего анализа использовали программный пакет Bionumerics 7.5
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