Molecular Screening of EEHV in Elephants

DNA was extracted from elephant blood samples using a DNA extraction kit (Machery-Nagel GmbH, Dauren, Germany), according to the manufacturer’s instructions. Specific primers, namely, pan-polymerase and terminase-specific primers, were used in PCR analysis for the screening of EEHV, as previously described [2 (link),18 (link),19 (link)]. Briefly, samples were screened for EEHV using PAN-EEHV POL primers 6710/6711 [10 (link)]. Then, the positive samples were confirmed for the EEHV subtype using terminase-specific primers for EEHV1 [2 (link)] and terminase-specific primers for EEHV3/4, as previously described [6 (link)] (Supplementary Table S1). The specific EEHV subtypes were further determined by sequencing, as has also been described previously [18 (link),19 (link)].