Creation of a hybridization probe for the diagnosis of classical swine fever by the FISH method

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Abstract

The article presents data creation of the DNA probe labeled with fluorescein-12-dUTP for its subsequent use in in-situ hybridization in cryotomic histological sections from samples of internal organs of pigs. This diagnostic method is especially relevant for the inapparate course of infection, which contributes to maintaining the viral load inside the herd without showing pronounced clinical signs. In addition, the method helps to identify the viral genetic material in the pathological samples during the post-viremia period. In this article the stages of creating a probe consisting of 173 pairs of nucleotide bases corresponding to the site of the glycoprotein E2 gene of the classical swine fever virus are shown. The final DNA probe was tested on tonsils from pigs infected with the CSF virus as a positive control for the DNA probe and as a negative control for the DNA probe is tonsils from healthy pigs not vaccinated against CSF were used. Finally, we obtained positive results of using the DNA probe, reflecting its high specificity and selectivity in the studied samples of positive control. The data obtained by us formed the basis for our further research in the field of infectious pathology of farm animals.

About the authors

A. K Komina

Federal State Budget Scientific Institution �Federal Scientific Centre VIEV�

V. V Stafford

Federal State Budget Scientific Institution �Federal Scientific Centre VIEV�

Email: stafford.v.v@gmail.com

A. M Gulyukin

Federal State Budget Scientific Institution �Federal Scientific Centre VIEV�

References

  1. Величко Н.В., Рабочая Д.Е., Макеева А.С., Пиневич А.В. Таксономическое разнообразие цианобактерий в антарктических почвенных сообществах, выявленное методом флуоресцентной in situ гибридизации. В сб.: Микроорганизмы и плодородие почвы. Мат. I Всеросс. науч.-практ. конф. с межд. уч., посвящ. 90-летию со дня рождения профессора Е.М. Панкратовой. 2022. С. 27-30.
  2. Каракулов Р.К., Кайдарова Д.Р., Душимова З.Д. и др. Дифференцированный подход к терапии диффузной В-крупноклеточной лимфомы с учетом статуса с-MYC и BCL2. Онкогематология. 2022. Т.17. №1. С. 37-42. doi: 10.17650/1818-8346-2022-17-1-37-42
  3. Шкаврова Т.Г., Михайлова Г.Ф., Цепенко В.В., Голуб Е.В. База данных частоты стабильных аберраций хромосом у детей, проживающих на радиоактивно-загрязненных территориях после чернобыльской аварии. Свидетельство о регистрации базы данных 2022620536, 15.03.2022. Заявка № 2022620389 от 09.03.2022.
  4. Choi C., Chae C. Localization of classical swine fever virus in male gonads during subclinical infection. J Gen Virol. 2002 Nov; 83(Pt 11):2717-2721. doi: 10.1099/0022-1317-83-11-2717.
  5. Choi C., Chae C. Detection of classical swine fever virus in the ovaries of experimentally infected sows. J Comp Pathol. 2003 Jan; 128(1):60-6. doi: 10.1053/jcpa.2002.0607.
  6. Fan Y-H, Lin Y-L, Hwang Y-C. et al. T-cell factor-4 and MHC upregulation in pigs receiving a live attenuated classical swine fever virus (CSFV) vaccine strain with interferon-gamma adjuvant. Vet J. 216. Р. 148-56. doi: 10.1016/j.tvjl.2016.07.009.
  7. Ha S.-K., Choi C., Chae C. Development of an optimized protocol for the detection of classical swine fever virus in formalin-fixed, paraffin-embedded tissues by seminested reverse transcription-polymerase chain reaction and comparison with in situ hybridization. Research in Veterinary Science. 2004. 77. Р. 163-169
  8. Nagarajan K., Saikumar G. Fluorescent in-situ hybridization technique for the detection and localization of classical swine fever virus in infected tissues. Veterinarski Arhiv. 82 (5). 2012. P. 495-504.
  9. URL: https://www.oie.int/en/what-we-do/standards/codes-and-manuals/terrestrial-manual-online-access
  10. Zhang Q., Xu Lu, Zhang Y. et al. A novel ViewRNA in situ hybridization method for the detection of the dynamic distribution of Classical Swine Fever Virus RNA in PK15 cells. Virology Journal. 2017. 14(1):81. doi: 10.1186/s12985-017-0734-4

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