Использование ПЦР и секвенирования для выявления и анализа генома коронавируса крупного рогатого скота
Диссертация
Применение методов молекулярной биологии, таких как ПЦР и секвенирование, открывает широкие возможности для выявления и изучения инфекционных агентов. От традиционных методов диагностики ПЦР отличает высокая аналитическая чувствительность и специфичность, что позволяют установить присутствие вируса в исследуемой пробе даже по фрагменту генома. Секвенирование амплифицированного в ПЦР участка… Читать ещё >
Содержание
- 1. ВВЕДЕНИЕ
- 2. ОБЗОР ЛИТЕРАТУРЫ
- 2. 1. Коронавирусная диарея новорожденных телят
- 2. 1. 1. Эпизоотология
- 2. 1. 2. Клинические признаки
- 2. 1. 3. Патогенез и патологоанатомические изменения
- 2. 1. 4. Диагностика
- 2. 1. 5. Специфическая профилактика
- 2. 2. Коронавирусная инфекция дыхательных путей
- 2. 3. «Зимняя дизентерия»
- 2. 4. Характеристика семейства Согопаутёае
- 2. 5. Структура вириона и физико-химические свойства коронавируса крупного рогатого скота
- 2. 6. Геном коронавируса крупного рогатого скота
- 2. 7. Основные структурные белки коронавируса крупного рогатого скота
- 2. 7. 1. Нуклеокапсидный протеин
- 2. 7. 2. Интегральный мембранный протеин
- 2. 7. 3. Спайковый гликопротеин
- 2. 7. 4. Гемагглютинин-эстеразный гликопротеин
- 2. 8. Репликация коронавируса крупного рогатого скота
- 2. 1. Коронавирусная диарея новорожденных телят
- 3. СОБСТВЕННЫЕ ИССЛЕДОВАНИЯ
- МАТЕРИАЛЫ И МЕТОДЫ
- 3. 1. Материалы
- 3. 1. 1. Штаммы вируса
- 3. 1. 2. Оборудование
- 3. 1. 3. Растворы и реактивы
- 3. 2. Методы
- 3. 2. 1. Выделение суммарной РНК
- 3. 2. 2. Реакция обратной транскрипции
- 3. 2. 3. Амплификация кДНК
- 3. 2. 4. Вариант ПЦР «continuous»
- 3. 2. 5. Очистка продуктов ПЦР
- 3. 2. 6. Реакция секвенирования
- 3. 2. 7. Компьютерный анализ данных
- 3. 1. Материалы
- 4. 1. Разработка и применение метода выявления и штаммовой дифференциации коронавируса крупного рогатого скота с помощью
- 4. 1. 1. Исследование профиля вариабельности НЕ гена
- 4. 1. 2. Выбор первичной структуры праймеров
- 4. 1. 3. Оптимизация условий постановки ПЦР и секвенирования
- 4. 1. 4. Апробация метода и анализ полученных данных
- 4. 2. Разработка и применение метода выявления и штаммовой дифференциации коронавируса крупного рогатого скота с помощью
- 4. 2. 1. Исследование профиля вариабельности S гена
- 4. 2. 2. Выбор праймеров, условий постановки ПЦР и секвенирования
- 4. 2. 3. Апробация метода и анализ полученных данных
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