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Báo cáo khoa học: "Development of a monoclonal antibody-based co-agglutination test to detect enterotoxigenic Escherichia coli isolated from diarrheic neonatal calves"

Tuyển tập các báo cáo nghiên cứu khoa học quốc tế về bệnh thú y đề tài: Development of a monoclonal antibody-based co-agglutination test to detect enterotoxigenic Escherichia coli isolated from diarrheic neonatal calves | J. Vet. Set. 2007 8 1 57-64 JOURNAL OF Veterinary Science Development of a monoclonal antibody-based co-agglutination test to detect enterotoxigenic Escherichia coli isolated from diarrheic neonatal calves Brajesh c. Varshney N.M. Ponnanna2 Pranati A. Sarkar2 Pragma Rehman2 Jigar H. Shah2 dntas Biopharmaceuticals Ltd. Plot No. 423 P A GIDC Sarkhej-Bavla Highway Moraiya Ahmedabad-382 210 Gujarat India 2R D Biotechnology National Daily Development Board Anand-388 001 Gujarat India Escherichia coll E. coli strains were collected from young diarrheic calves in farms and field. Strains that expressed the K99 F5 antigen were identified by agglutination tests using reference antibodies to K99 antigen and electron microscopy. The K99 antigen from a selected field strain SAR-14 was heat-extracted and fractionated on a Sepharose CL-4B column. Further purification was carried out by sodium deoxycholate treatment and or ion-exchange chromatography. Monoclonal antibodies to purified K99 antigen were produced by the hybridoma technique and a specific clone NEK99-5.6.12 was selected for propagation in tissue culture. The antibodies thus obtained were affinity-purified characterized and coated onto Giemsa-stained Cowan-I strain of Staphylococcus aureus S. aureus . The antibody-coated A aureus were used in a coagglutination test to detect K99 E. coll isolated from feces of diarrheic calves. The specificity of the test was validated against reference monoclonal antibodies used in co-agglutination tests as well as in ELISA. Specificity of the monoclonal antibodies was also tested against various Gram negative bacteria. The developed antibodies specifically detected purified K99 antigen in immunoblots as well as K99 E. coll in ELISA and co-agglutination tests. The co-agglutination test was specific and convenient for large-scale screening of K99 E. coll isolates. Key words co-agglutination diarrhea Escherichia coll K99 antigen monoclonal antibodies Introduction Diarrhea-causing .