Generation and validation of new quantitative real time PCR assays to detect elephant endotheliotropic herpesviruses 1A, 1B, and 4

Elephant endotheliotropic herpesviruses (EEHVs) can cause fatal hemorrhagic disease in Asian and African elephants. There are quantitative real time PCR (qPCR) tests that can detect seven known EEHVs (1A, 1B, 2-6) in mucosal secretions, tissue isolates, and blood samples. However, current qPCR tests are unable to distinguish between EEHV 1A and 1B or 3 and 4. To address these inadequacies, new qPCR assays were generated and validated to specifically detect EEHV 1A, 1B, and 4. Each assay demonstrated robust efficiency, a broad linear range, and low intra- and inter-assay variability. Each also proved to be specific for its EEHV target when tested against known banked samples from past EEHV cases. The EEHV1A and 1B assays were then used to characterize an eight-week, low level EEHV1 viremic event in a young Asian elephant. These new tests will allow veterinarians and researchers to pinpoint the specific species causing infection more rapidly. They will also allow veterinarians and elephant keepers to better characterize the EEHV status of each animal within their herd leading to more informed management strategies.