Разработка средств и методов лабораторной диагностики пограничной болезни овец тема диссертации и автореферата по ВАК РФ 16.00.03, кандидат ветеринарных наук Луговцев, Владимир Юрьевич

• Луговцев, Владимир Юрьевич
• кандидат ветеринарных наук
• 2000

Оглавление диссертации кандидат ветеринарных наук Луговцев, Владимир Юрьевич

1 Содержание

2 Список сокращений

3 Введение

3.1. Актуальность темы

3.2. Цели и задачи исследования

3.3. Научная новизна

3.4. Практическая значимость

3.5. Основные положения диссертационной работы, выносимые на защиту

3.6. Апробация работы

3.7. Публикации

4 Обзор литературы

4.1. Характеристика по^ганичной болезни овец

4.1.1. Историческая справка и экономическая значимость болезни

4.1.2. Эпизоотология ПБО

4.1.3. Патогенез ПБО

4.1.4. Клиническое проявление ПБО

4.1.5. Патологоанатомическая картина при ПБО

4.1.6. Иммунитет при ПБО

4.1.7. Профилактика и меры борьбы при ПБО

4.2. Характеристика возбудителя ПБО

4.2.1 .Таксономическое положение вируса ПБО

4.2.2. Физико-химические свойства вируса ПБО

4.2.3. Генетическая структура вируса ПБО

4.2.4. Молекулярная биология вируса ПБО

4.2.5. Антигенное и генетическое родство вируса ПБО с другими пестивирусами

4.3. Диагностика ПБО

4.3.1. Вирусологические методы исследования

4.3.1.1. Выделение вируса ПБО в культуре клеток

4.3.1.2. Методы обнаружения антигена вируса ПБО иммуноферментным анализом

4.3.1.3. Обнаружение и идентификация вирусной РНК

4.3.2. Методы выявления специфических антител

4.3.2.1. Реакция нейтрализации

4.3.2.2. Методы на основе иммуноферментного анализа 31 4.3.3. Дифференциальная диагностика

Заключение диссертации по теме «Ветеринарная эпизоотология, микология с микотоксикологией и иммунология», Луговцев, Владимир Юрьевич

7 Выводы

1. Изучены некоторые параметры репродукции вируса ПБО в первичных и перевиваемых культурах клеток. Наиболее высокой чувствительностью обладают первичные культуры клеток ТЯ и ПЯ, в которых он накапливается в титре 4,3±0,15 lg ККИД50/см3 в течение 96-120 часов.

2. Изучена инфекционность вируса ПБО на овцах романовской породы. Установлена трансплацентарная передача вируса, сопровождающаяся образованием специфических вируснейтрализующих антител в организме плода, при инфицировании овцематки на поздней стадии суягности.

3. Показано, что уровнь ПКА лейкоцитов периферической крови, инфицированных пестивирусами in vitro или in vivo, не может служить объективным критерием дифференциации пестивирусов.

4. Разработаны схемы иммунизации, обеспечивающие получение на овцах специфических сывороток к вирусу ПБО с активностью в реакции нейтрализации 1:512-1:1024, и полипестивирусных сывороток на свиньях, которые могут быть использованы в диагностических тестах для типирования вируса ПБО в реакции нейтрализации и обнаружения специфического вирусного антигена в культуре клеток в реакции иммунофлюоресценции.

5. Разработана методика обнаружения и идентификации РНК вируса ПБО на основе полимеразной цепной реакции.

6. Разработана схема лабораторной диагностики ПБО, включающая отработанные методики выделения вируса в культуре клеток и его идентификации в реакции иммунофлюоресценции; перекрестной реакции нейтрализации -для серологического типирования вируса ПБО; полимеразной цепной реакции -для обнаружения и дифференциации РНК вируса ПБО и реакции нейтрализации - для выявления специфических антител.

7. При исследовании полевых сывороток крови овец, полученных из разных регионов России, в 18,2% случаев выявлены специфические антитела к пестивирусам жвачных в титре от 1:8 до 1:128, что свидетельствует о возможной циркуляции пестивирусов в популяциях овец. В исследованных пробах сывороток крови коз антител не обнаружено.

Практические предложения 132

8 Практические предложения

На основе разработанных средств, методов и схемы лабораторной диагностики ПБО предложены "Методические указания по лабораторной диагностике пограничной болезни овец", утвержденные директором ВНИИВВиМ, внедренные в научно-исследовательскую и диагностическую работу в институте и рекомендуемые для широкого использования в практике диагностических исследований.

Благодарности

Отдельные этапы комплексных исследований выполнены совместно с кандидатом ветеринарных наук В.В. Куринновым; доктором биологических наук С.Ж. Цыбановым; кандидатом биологических наук Ю.О. Селяниновым; кандидатом биологических наук Е.К. Сенечкиной; кандидатом биологических наук Н.Г. Шубиной; кандидатом биологических наук Б.В. Новиковым; кандидатом биологических наук JI.B. Шевченко; кандидатом биологических наук Н.С. Неверовской; кандидатом биологических наук С.И. Сидоровым; научным сотрудником Л.И. Анисимовой, младшим научным сотрудником Н.Г. Афанасьевой, за что автор выражает им глубокую признательность.

Искреннюю благодарность автор приносит коллективу лаборатории Диагностики за оказанную помощь при выполнении данной работы.

Автор благодарен профессору H.A. Лагуткину, доктору биологических наук H.A. Власову, кандидату биологических наук С.Г. Юркову (ВНИИВВиМ), доктору ветеринарных наук С. А. Жидкову (ВИЭВ) за ценные консультации.

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