Production and identification of IgY against excretory-secretory antigens of Trichinella spiralis muscle larvae.

To produce and purify egg yolk immunoglobulin(IgY)against excretory-secretory(ES)antigens of T.spiralis muscle larvae and assay its titer and sensitivity in antigens detection,twenty-four-week old hens were intramuscularly immunized with ES antigens of T.spiralis muscle larvae for 4 times.Four hens were firstly immunized with 500 μg of ES antigens and subsequent immunization was performed at 10-days intervals with 250 μg of ES antigens.IgY was extracted by water-dilution method from eggs of non-immunized hens and immunized hens 42 days after the first immunization.The protein concentration of IgY was measured by Bradford method,and IgY was analyzed by SDS-PAGE,Western blot and IFAT.ELISA was used to assay the titer of IgY and its sensitivity to detect antigens.The results showed that about 70 mg of IgY was extracted from each egg.SDS-PAGE showed IgY contained two major protein bands with molecular weight of 67 kDa and 23 kDa.The results of Western blot and IFAT demonstrated that somatic and ES antigens of T.spiralis muscle larvae could be recognized by IgY.The purified IgY antibody titer was 1∶107.The sensitivity of IgY-sandwich ELISA to detect antigens was 1.17ng/mL.It's demonstrated that the IgY to ES antigens of T.spiralis muscle larvae had high antibody titer and sensitivity.