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Разработка тест системы для диагностики африканской чумы свиней методом ПЦР-РВ

выпускная квалификационная работа бакалавра

Разработка тест системы для диагностики африканской чумы свиней методом ПЦР-РВ

Abstract

На 68 страниц, 10 рисунков, 9 таблиц.Тема выпускной квалификационной работы: «Разработка тест-системы для диагностики вируса африканской чумы свиней методом ПЦР-РВ».Целью данной работы является создание диагностической тест-системы, основанной на методе полимеразной цепной реакции в режиме реального времени, для идентификации вируса африканской чумы свиней.Задачи, которые решались в ходе исследования:Подбор матрицы-мишени insilico.Отбор изолятов вируса из National Center for Biotechnology Information, США (NCBI).Определение консенсусной последовательности гена B646L и подбор праймеров к ней.Подбор последовательности гена B646L для положительного контрольного образца (ПКО), подбор последовательности гена Цитохрома В для контроля взятия материала (КВМ) и соответствующих праймеров.Олигонуклеотидный синтез ранее подобранных праймеров.Сбор тест-системы и проверка ее работоспособности.В качестве основных методов исследования выбраны олигонуклеотидный синтез, секвенирование, клонирование бактерий, анализы генетических последовательностей insilico, ПЦРв режиме реального времени.Ген B646L, кодирующий основной капсидный белок р72, вируса африканской чумы свиней является наиболее консервативной последовательностью. На основе фрагмента последовательности выбранного гена сконструирована диагностическая ПЦР-РВ тест-система.Данная тест-система может быть использована для диагностики вируса африканской чумы свиней в образцах, представляющих из себя буккальный эпителий, кровь, лимфу, ткани пораженных органов.

68 pages, 10 figures, 9 tablesThe subject is «Design test-system for diagnosing African swine fever virus by method of PCR-RT».The purpose of this scientific work is to design a diagnostic test-system based on the real-time polymerase chain reaction method for the identification of the African swine fever virus.Tasks that were solved in the course of the study:• Selection of the target matrix in silico.• Selection of virus isolates from the National Center for Biotechnology Information, USA (NCBI).• Determination of the consensus sequence of the B646L gene and selection of primers for it.• Sequencing of the B646L gene for the positive control sample (PQS), sequencing of the Cytochrome B gene for sampling control (CVM) and appropriate primers.• Oligonucleotide synthesis of previously selected primers.• Collecting the test system and checking its performance.Oligonucleotide synthesis, sequencing, bacterial cloning, in silico genetic sequence analyzes, real-time PCR were chosen as the main research methods.The B646L gene encoding the p72 major capsid protein of the African swine fever virus is the most conserved sequence. A diagnostic PCR-RT test-system was built on the basis of a fragment from this gene.This test-system can be used to diagnose the African swine fever virus in samples, which are buccal epithelium, blood, lymph, tissues of the affected organs.

Keywords

диагностика., полимеразная цепная реакция (ПЦР), diagnosis, polimerase chain reaction (PCR), вирус африканской чумы свиней (ВАЧС), african swine fever virus (ASFV)

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selected citations
These citations are derived from selected sources.
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
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