EHDV Serotyping by One-Step qRT-PCR

The samples that were positive during the initial EHDV detection were then tested for serotypes 1, 2, and 6. The reactions were performed in technical triplicates using the Superscript III/ Platinum Taq One-Step qRT-PCR Kit (Invitrogen, Waltham, MA, USA). The primers and probes (LGC Biosearch Technologies, Petaluma, CA, USA) were specific to the VP2, seg-2 region of each virus serotype [24 (link)] (Supplementary Table S1). The samples were amplified on a Bio-Rad CFX OPUS real-time PCR machine under the following conditions: 55 °C for 30 min, 95 °C for 10 min, and 50 cycles at 95 °C for 30 s and 60 °C for 1 min. The data were analyzed using Bio-Rad CFX Maestro qPCR software.