Development of an immunochromatographic strip test for antigen detection of elephant endotheliotropic herpesvirus in Asian elephants (Elephas maximus)

Thunyamas Guntawang; Tidaratt Sittisak; Saralee Srivorakul; Kornravee Photichai; Pisinee Aiumurai; Chatchote Thitaram; Nattawooti Sthitmatee; Wei-Li Hsu; Nitat Sookrung; Kidsadagon Pringproa

doi:10.1016/j.jviromet.2022.114627

Development of an immunochromatographic strip test for antigen detection of elephant endotheliotropic herpesvirus in Asian elephants (Elephas maximus)

J Virol Methods. 2023 Jan:311:114627. doi: 10.1016/j.jviromet.2022.114627. Epub 2022 Sep 30.

Affiliations

¹ Department of Veterinary Biosciences and Veterinary Public Health, Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai 50100, Thailand.
² Veterinary Diagnostic Center, Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai 50100, Thailand.
³ Biomedical Research Incubator Unit, Department of Research, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand.
⁴ Center of Elephant and Wildlife Health, Chiang Mai University, Chiang Mai 50100, Thailand; Department of Companion Animals and Wildlife Clinics, Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai 50100, Thailand.
⁵ Graduate Institute of Microbiology and Public Health, College of Veterinary Medicine, National Chung Hsing University, Taichung 402, Taiwan.
⁶ Department of Veterinary Biosciences and Veterinary Public Health, Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai 50100, Thailand; Center of Elephant and Wildlife Health, Chiang Mai University, Chiang Mai 50100, Thailand. Electronic address: kidsadagon.p@cmu.ac.th.

PMID: 36191664
DOI: 10.1016/j.jviromet.2022.114627

Abstract

Elephant endotheliotropic herpesvirus (EEHV) is the causative agent of EEHV-hemorrhagic disease (EEHV-HD) in elephants worldwide. This disease is highly virulent and a predominant cause of fatalities in young Asian elephants. Rapid diagnosis and aggressive therapies have been determined to be a key strategy in the successful treatment of this disease. Herein, we have developed the immunochromatographic strip test for EEHV detection. Accordingly, 31.2 kDa of partial EEHV DNA polymerase (DNApol) protein was expressed in Escherichia coli and used to generate rabbit polyclonal anti-EEHV DNApol antibodies. These were then used to develop an ICS test for EEHV antigen detection using the double-antibody sandwich colloidal gold method. Anti-EEHV DNApol antibodies conjugated with 40 nm colloidal gold solution were used as a detector, while rabbit anti-EEHV DNApol and goat anti-rabbit IgG antibodies immobilized on the nitrocellulose membrane were used as the test and control lines, respectively. The test had a detection limit of 1.25 × 10⁵ viral genome copies (vgc)/mL of EEHV obtained from blood samples. Moreover, no specialized equipment or laboratory infrastructure was required in the administration of this test. This developed ICS test for EEHV antigen detection can be used in field application for the rapid detection of EEHV in resource-limited environments.

Keywords: DNA polymerase; EEHV; EEHV-hemorrhagic disease; Immunochromatographic strip test.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Antigens, Viral
Elephants*
Gold Colloid
Herpesviridae Infections* / diagnosis
Herpesviridae Infections* / veterinary
Herpesviridae* / genetics
Rabbits

Substances

Antigens, Viral
Gold Colloid