A lytic phage for Pseudomonas aeruginosa belongs to the Myoviridea family was isolated from urine for use in therapeutics. Pair of hepatitis C virus (HCV) primers highlighted segments on the genome of this phage. The sequence of these PCR products as well as the possible serological cross reactivity/relationship between HCV and the phage were investigated. One hundred HCV positive human sera were analyzed by ELISA. Ninety six well plates were coated with multiple epitopes of HCV proteins (Kit), phage and Pseudomonas cells. Initially the positive and negative control sera supplied in the test kit were used to evaluate the cross reactivity between the phage and anti-HCV antibodies. The results suggested a value over than 0.105 for a HCV positive reaction. Of the 100 HCV positive sera tested, sixty five and thirty percent showed cross reaction with phage lysate and Pseudomonas aeruginosa, respectively. High HCV antibody titer correlated to high cut off value for phage cross reaction, whereas no such correlation existed between HCV antibody titer and Pseudomonas cross reaction. The PCR products were sequenced and aligned with the HCV genome of H77. Sequence homology was detected in the 5', 3' UTRs and NS3 regions. Further these products showed similarity with HIV-1 Env, Pol & 3'LTR regions as well.