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. 2023 Mar 30;85(4):459-462.
doi: 10.1292/jvms.22-0578. Epub 2023 Mar 15.

Direct detection of elephant endotheliotropic herpesvirus 1 (EEHV1) DNA in heparinized plasma by loop-mediated isothermal amplification

Kazuya Takehana  1 , Keita Matsuno  2   3   4
Affiliations

Direct detection of elephant endotheliotropic herpesvirus 1 (EEHV1) DNA in heparinized plasma by loop-mediated isothermal amplification

Kazuya Takehana et al. J Vet Med Sci. .

Abstract

Elephant endotheliotropic herpesvirus (EEHV) causes a fatal hemorrhagic disease and is a significant cause of mortality in juvenile Asian elephants. A loop-mediated isothermal amplification (LAMP) method was developed to rapidly diagnose EEHV viremia. However, extracting DNA from whole blood samples to perform LAMP hampers diagnosis in a field setting. Here, we established the Direct-LAMP method, using heparinized plasma without extracting the DNA to speed up and simplify the test. EEHV-positive specimens were tested using the Direct-LAMP. The detection limit was calculated to be 101.3 copies/μL using the mimetic samples, which was almost identical to the value determined in LAMP in which DNA was extracted. Hence, the Direct-LAMP provided a more rapid diagnosis to save, which could prevent elephant deaths.

Keywords: Asian elephants (Elephas maximus); direct detection; elephant endotheliotropic herpes virus (EEHV); heparinized plasma; loop-mediated isothermal amplification (LAMP).

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Conflict of interest statement

No author has any conflict of interest.

References

    1. Boonprasert K, Punyapornwithaya V, Tankaew P, Angkawanish T, Sriphiboon S, Titharam C, Brown JL, Somgird C. 2019. Survival analysis of confirmed elephant endotheliotropic herpes virus cases in Thailand from 2006–2018. PLoS One 14: e0219288. doi: 10.1371/journal.pone.0219288 - DOI - PMC - PubMed
    1. Dastjerdi A, Seilern-Moy K, Darpel K, Steinbach F, Molenaar F. 2016. Surviving and fatal elephant endotheliotropic herpesvirus-1A infections in juvenile Asian elephants-lessons learned and recommendations on anti-herpesviral therapy. BMC Vet Res 12: 178. doi: 10.1186/s12917-016-0806-5 - DOI - PMC - PubMed
    1. Fuery A, Tan J, Peng R, Flanagan JP, Tocidlowski ME, Howard LL, Ling PD. 2016. Clinical infection of two captive Asian elephants (Elephas maximus) with elephant endotheliotropic herpesvirus 1B. J Zoo Wildl Med 47: 319–324. doi: 10.1638/2015-0074.1 - DOI - PubMed
    1. Goto M, Honda E, Ogura A, Nomoto A, Hanaki K. 2009. Colorimetric detection of loop-mediated isothermal amplification reaction by using hydroxy naphthol blue. Biotechniques 46: 167–172. doi: 10.2144/000113072 - DOI - PubMed
    1. Guntawang T, Sittisak T, Srivorakul S, Photichai K, Aiumurai P, Thitaram C, Sthitmatee N, Hsu WL, Sookrung N, Pringproa K. 2023. Development of an immunochromatographic strip test for antigen detection of elephant endotheliotropic herpesvirus in Asian elephants (Elephas maximus). J Virol Methods 311: 114627. doi: 10.1016/j.jviromet.2022.114627 - DOI - PubMed

Supplementary concepts