Development and validation of quantitative real-time polymerase chain reaction assays to detect elephant endotheliotropic herpesviruses-2, 3, 4, 5, and 6

► Real-time PCR assays were developed for EEHV2, 3, 4, 5, and 6. ► The new EEHV assays are sensitive and specific. ► The assays are useful for determining EEHV prevalence in in situ and ex situ elephants. ► The assays will be useful for diagnosing suspected cases of EEHV-associated illness. ► The assays will be useful for managing treatment of elephants with EEHV-induced illness.