Expression of ORF2 protein of TTV for development of EIA system

Methods ORF2 region of TTV genotype1 (JA20 isolate) was amplified using gene specific primers and cloned in pET19b expression vector for protein expression. Expressed protein was confirmed by western blotting using anti-His antibodies against N-terminal 6XHis tag of expressed protein. Antigenicity of ORF2 was determined by western blot, detecting anti-TTV antibodies in 50 human sera of which, 10 were healthy control negative for TTV-DNA, 10 liver disease cases negative for TTV-DNA and 30 liver disease cases that were positive for TTV-DNA.