DNA Extraction
Skin scabs from pathological tissues were centrifuged at 10,000 g for 10 min at +4 °C. The centrifuged scabs was used for viral genomic DNA extraction using Blood-Animal-Plant DNA Preparation kit (Jena Biosciences®, Germany). A total of 300 µl of lysis buffer was added to 200 µl of tissue homogenate and 2 µl RNAse A inhibitor was added and vortexed for 30 seconds, then 8µl of proteinase K was added to digest the tissue and placed on heating block at 60oC for 30 minutes. This was cooled for 5mins and 300 µl binding buffer was added and centrifuged at 10,000g for 5mins. The supernatant was decanted into a spin column and washed twice with wash buffer. Fifty microliter of viral DNA was eluted into a new eppendorf tube, labeled and kept at -4oC briefly before PCR.