Development Of Indirect Enzyme-Linked Immunosorbent Assay For Detecting Antibody Against Swine Chlamydophila Abortus

Chlamydophila abortus(CAB) was one of the important pathogens causing abortion of sow,cow and ewe and pregnant women.CAB infection in intensive pig farms must be paid enough attention.Development of rapid and accurate diagnostic methods is the essential measure for the prevention and control of this disease.In this study,the primers were designed according to the polymorphic outer membrane protein-90(pomp90) gene sequence of CAB published in Genbank and four different fragments of pomp90 gene were obtained by PCR.The recombinant plasmids were constructed and expressed in E.coli BL21,and the recombinant POMP90 proteins were purified and renaturated. Applicabilities of the high purity and immunologic recombinant proteins were explored and the indirect ELISA methods for detecting the CAB antibody in sera of pigs were developed.1.Four different fragments of pomp90 gene were amplified with the specific primers and the total DNA template of CAB strain CP/12.The obtained genes were identified by sequencing after linked to the the prokaryotic expression vector pGEX-KG.2.The recombinant plasmids were expressed in the E.coli BL21,The four purified fusion POMP-GST protein showed good immune reactivity detected by Western-blot3.The indirect ELISA methods for the detection of the CAB antibody were established using the four purified recombinant proteins as coating antigens after the conditions were optimized.Then the mothods were clinically used to detect the serum antibody.The results showed that the methods developed with the four recombinant POMPs were sensitive,specific,accurate,and to overcome the shortcomings of traditional IHA,such as low sensitivity,subjective judgments,and so on.In this paper,the recombinant expression plasmids were constructed which were expressed in E.coli BE21.Then coated with the purified protein,the indirect ELISA methods were established for detecting the CAB antibodies in the swine sera.This study was the basis for the further research on diagnosis of swine Chlamydophila abortus infection.