Mature flukes of Clonorchis sinensis were recovered from the gall bladder and bile ducts of infected cats. Flukes were washed with sterilized normal saline, passed through a serial sterile procedure and incubated at 37°C in a 5% CO2 atmosphere. After one week, flukes were changed to fresh culture medium and prepared for heat shock treatment. Treatment conditions are described as follows: 4°C for 120 min, followed by 25, 37 or 42°C each for 60 min, and 45°C for 30 min. SDS-PAGE of C. sinensis adult fluke extracts revealed a complex pattern of 34 polypeptide components which ranged from 8.5 to 112 kDa. When temperature rose to 42°C, two significant C. sinensis heat shock proteins (75 and 82 kDa) appeared. Results of electrotransfer and immunoblotting showed that the sites of heat-induced antigens can be recognized when using a sera pool of 25 clonorchiasis patients.
Chinese liver fluke, Stress protein
