Development of in situ hybridization for detection of elephant endotheliotropic herpesvirus in Asian elephants

Varankpicha Kochakul; Kittikorn Boonsri; Saruda Tiwananthagorn; Chalermchart Somgird; Chatchote Thitaram; Kidsadagon Pringproa

doi:10.1177/1040638718773810

Development of in situ hybridization for detection of elephant endotheliotropic herpesvirus in Asian elephants

J Vet Diagn Invest. 2018 Jul;30(4):628-632. doi: 10.1177/1040638718773810. Epub 2018 May 7.

Authors

Varankpicha Kochakul^{1

2

3

4}, Kittikorn Boonsri^{1

2

3

4}, Saruda Tiwananthagorn^{1

2

3

4}, Chalermchart Somgird^{1

2

3

4}, Chatchote Thitaram^{1

2

3

4}, Kidsadagon Pringproa^{1

2

3

4}

Affiliations

¹ Veterinary Diagnostic Laboratory (Kochakul, Boonsri), Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai, Thailand.
² Department of Veterinary Biosciences and Veterinary Public Health (Tiwananthagorn, Pringproa), Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai, Thailand.
³ Department of Companion Animals and Wildlife Clinics (Somgird, Thitaram), Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai, Thailand.
⁴ Center of Excellence in Elephant Research and Education (Somgird, Thitaram, Pringproa), Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai, Thailand.

Abstract

Elephant endotheliotropic herpesvirus (EEHV) is one of the most important viral infectious diseases affecting the elephant population worldwide, especially juveniles and young adults. We developed a chromogenic in situ hybridization (ISH) test for detection of EEHV in Asian elephants ( Elephas maximus). Digoxigenin (DIG) DNA probes from the polymerase and terminase genes of EEHV were synthesized using a PCR DIG-labeling method, and detection of hybridized probe to target EEHV DNA was carried out by anti-DIG immunolabeling. Distribution of EEHV-1A and EEHV-4 genomes was found to be prominent in mononuclear phagocytic cells of spleen and endothelial cells of visceral organs. ISH enables the detection of EEHV infection and has applications in understanding pathogenesis of EEHV in Asian elephants.

Keywords: Asian elephants; elephant endotheliotropic herpesvirus; in situ hybridization.

MeSH terms

Animals
Elephants / virology*
Genome, Viral
Herpesviridae / genetics
Herpesviridae Infections / diagnosis
Herpesviridae Infections / veterinary*
Herpesviridae Infections / virology
In Situ Hybridization / methods
In Situ Hybridization / veterinary*
Polymerase Chain Reaction / veterinary