Оптимизация условий постановки ИФА для диагностики вируса ящура типа А
DOI:
https://doi.org/10.26577/eb-2019-1-1411Аннотация
Путем концентрирования ПЭГ-6000 и очистки хлороформом штамма «Таласский» вируса ящура типа А приготовлены очищенные антигены, активность которых составила в РСК 1:32- 1:64. Для получения диагностических сывороток антигены подвергались дополнительной очистке через 30% сахарозную подушку при 35000 об/мин в течение 2 часов. После очистки активность антигена составила 1:16-1:32. Используя очищенный антиген вируса ящура типа А, были получены антисыворотки на овцах, свиньях и кроликах, с активностью 1:8-1:64. Из антисывороток методом Кона выделен иммуноглобулин, активность которого в РДП составила 1:32, а концентрация белка в пределах от 41,84 мг/мл до 60,2 мг/мл. Используя периодатный метод M.B.Wilson, P.K.Nakane на основе специфического свиного глобулина, приготовлен конъюгат для оптимизации условий постановки ИФА. Приготовленные диагностические препараты использованы для оптимизации условий постановки ИФА для диагностики ящура типа А. Оптимизированный вариант ИФА оказался чувствительным и эффективным при исследовании гетерогенных и гомогенных биоматериалов. Ключевые слова: ящур типа А, иммуноферметный анализ, иммуноглобулин, иммунизация, антиген.
Библиографические ссылки
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assay (ELISA) for type identification of foot-and-mouth-disease (FMD) virus in direct field materials” // Indian Journal of
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24 Kistler P., Nitschman H. Production of proteins from plasma // Vox. Sang., 1962. – V.7. – P. 414– 424.
25 Мищенко В.А., Шажко Ж.А., Базаров М.А., Конюшкина Т.Б., Смирнов А.Б., Атаев Э.А. Сравнительная оценка
методов конъюгации антител к вирусу ящура с пероксидазой хрена // Акт. пробл. вет. вирусологии. Тез. докл. науч. конф.
ВНИЯИ, – Владимир, 1987.- 2.- С. 49 – 50.
26 Щербаков А.В. Молекулярная эпизоотология ящура в России (филогенетический анализ российских изолятов
вируса ящура) // Ветеринария сегодня. 2015.-(3).- С. 30-36.
disease virus by real-time, fluorogenic reverse transcription polymerase chain reaction assay // J. Virol. Meth. – 2002.
– Vol. 105. – PР. 67-80.
2 Foot And Mouth Disease Virus/Manual of Standards for Diagnostic Tests and Vaccines – Paris: OIE.- 1996.- РР. 47 – 57.
3 Kitching R. P. Barnett P. V. Donaldson A. I. Mackay D. K. J. Foot and mouth disease. Manual of standards for diagnostic
tests and vaccines: Lists A and B diseases of mammals, birds and bees Paris: OIE.- 2001.- РР. 77-92.
4 Nagendrakumar S.B., Reddy G.S., Chandran D., Thiagarajan D., Rangarajan P.N., Srinivasan V.A. Molecular characterization
of foot-and-mouth disease virus type C of Indian origin // J. Clinical Microbiol. – 2005. – Vol. 43. N. 2. – PР. 966-969.
5 Have P., Lei J. C., Schjerning-Thiesen K. “An engyme linked immunosorbent assay (ELISA) for the primary diagnosis of
foot-and-mouth disease: characterization and comparison with complement fixation” // Acta Veterinaria Scandinavica. Supplementum.
– Vol. 25. – No. 2. – PP. 280–296. – 1984.
6 Ferris N. P., Dawson M. “Routine application of enzyme-linked immunosorbent assay in comparison with complement fixation
for the diagnosis of foot-and-mouth and swine vesicular diseases” // Veterinary Microbiology. – Vol. 16. – No. 3. – PP. 201–209.
– 1988.
7 Abu Elzein E. M. E., Crowther J. R. “Enzyme-labelled immunosorbent assay techniques in foot-and-mouth disease virus
research” // Journal of Hygiene. – Vol. 80. – No. 3. – PP. 391–399. – 1978.
8 Crowther J. R., Abu Elzein E. M. E. “Application of the enzyme linked immunosorbent assay to the detection and identification
of foot-and-mouth disease viruses” // Journal of Hygiene. – Vol. 83. – No. 3. – PP. 513–519. – 1979.
9 Ouldridge E. J., Barnett P., Rweyemamu M. M. “The relative efficiency of two ELISA techniques for the tritration of FMD
antigen” // Current Topics in Veterinary Medicine and Animal Science. – Vol. 22. – PP. 142–151. – 1982.
10 Abu Elzein E. M. E., J. R. Crowther. “The specific detection of foot-and-mouth disease virus whole particle antigen (140S)
by enzyme labelled immunosorbent assay” // Journal of Hygiene. – Vol. 83. – No. 1. – PP. 127–134. – 1979.
11 Rai A., Lahiri D. K. “A micro-enzyme-labelled immunosorbent assay (MICROELISA) for the detection of foot-and-mouth
disease virus antigen and antibody” // Acta Virologica. – Vol. 25. – No. 1. – PP. 49–52. – 1981.
12 Hamblin C., Armstrong R. M., Hedger R. S. “A rapid enzyme-linked immunosorbent assay for the detection of foot-andmouth
disease virus in epithelial tissues” // Veterinary Microbiology. – Vol. 9. – No. 5. – PP. 435–443. – 1984.
13 Ouldridge E. J., Barnett P. V., Hingley J., Rweyemamu M. M. “The differentiation of foot and mouth disease virus strains
using an indirect sandwich enzyme-linked immunosorbent assay saturation model” // Journal of Biological Standardization. – Vol.
12. – No. 4. – PP. 367–377. – 1984.
14 Roeder P. L., Le Blanc Smith P. M. “Detection and typing of foot-and-mouth disease virus by enzyme-linked immunosorbent
assay: a sensitive, rapid and reliable technique for primary diagnosis” // Research in Veterinary Science. – vol. 43. – No. 2. – PP.
225–232. – 1987.
15 Pattnaik B., Venkataramanan R. “Indirect enzyme linked immunosorbent assay for the detection of FMDV antigen” // Indian
Journal of Animal Sciences. – Vol. 59. – PP. 317–322. – 1989.
16 Pattnaik B., Venkataramanan R. “A sandwich enzyme linked immunosorbent assay for subtype analysis of foot-and-mouth
disease virus isolate” // Indian Journal of Animal Sciences. – Vol. 59. – PP. 1363–1368. – 1989.
17 Сюрин В.Н., Белоусова Р.В, Фомина Н.В. Диагностика вирусных болезней животных // М., "Агропромиздат"- 1991г.
– С. 434 – 463.
18 Paiba G. A., Anderson J., Paton D. J. “Validation of a foot-and-mouth disease antibody screening solid-phase competition
ELISA (SPCE)” // Journal of Virological Methods. – Vol. 115. – No. 2. – PP. 145–158. – 2004.
19 Bhattacharya S., Pattnaik B., Venkataramanan R. “Development and application of a sandwich enzyme-linked immunosorbent
assay (ELISA) for type identification of foot-and-mouth-disease (FMD) virus in direct field materials” // Indian Journal of
Animal Sciences. -Vol. 66. – No. 12. – PP. 1201–1209. – 1996.
20 Grubman M. J., Baxt B. “Foot-and-mouth disease” // Clinical Microbiology Reviews. – Vol. 17. – No. 2. – PP. 465–493. –
2004.
21 Мищенко В.А. Иммуноферментный метод в диагностике ящура // Вопр. вирусол. – 1991. – №4. – С. 341 -342.
22 Cohn E., Strong L., Hughes W. Fractionation of proteins from plasma //J. Amer. Chem. Soc.- 1946. – V.68. – 459 р.
23 Oncley J., Melin M., Richert D. The separation of the antibodies //J. Amer. Chem. Soc., 1949. – V.71. – PР. 541-550.
24 Kistler P., Nitschman H. Production of proteins from plasma // Vox. Sang., 1962. – V.7. – P. 414– 424.
25 Мищенко В.А., Шажко Ж.А., Базаров М.А., Конюшкина Т.Б., Смирнов А.Б., Атаев Э.А. Сравнительная оценка
методов конъюгации антител к вирусу ящура с пероксидазой хрена // Акт. пробл. вет. вирусологии. Тез. докл. науч. конф.
ВНИЯИ, – Владимир, 1987.- 2.- С. 49 – 50.
26 Щербаков А.В. Молекулярная эпизоотология ящура в России (филогенетический анализ российских изолятов
вируса ящура) // Ветеринария сегодня. 2015.-(3).- С. 30-36.
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Опубликован
2019-05-04
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БИОТЕХНОЛОГИЯ
