Journal of Veterinary Science

  • 제8권1호
  • /
  • Pages.57-64
  • /
  • 2007
  • /
  • 1229-845X(pISSN)
  • /
  • 1976-555X(eISSN)
대한수의학회 (The Korean Society of Veterinary Science)
권호 표지
DOI QR코드

DOI QR Code

Development of a monoclonal antibody-based co-agglutination test to detect enterotoxigenic Escherichia coli isolated from diarrheic neonatal calves

  • Varshney, Brajesh C. (Intas Biopharmaceuticals Ltd.) ;
  • Ponnanna, N.M. (R&D, Biotechnology, National Dairy Development Board) ;
  • Sarkar, Pranati A. (R&D, Biotechnology, National Dairy Development Board) ;
  • Rehman, Pragna (R&D, Biotechnology, National Dairy Development Board) ;
  • Shah, Jigar H. (R&D, Biotechnology, National Dairy Development Board)
  • 발행 : 20070300
⟨ 이전 논문 다음 논문 ⟩

초록

Escherichia coli (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 Giemsastained Cowan-I strain of Staphylococcus aureus (S. aureus). The antibody-coated S. aureus were used in a coagglutination test to detect K99+ E. coli 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. coli in ELISA and co-agglutination tests. The co-agglutination test was specific and convenient for large-scale screening of K99+ E. coli isolates.

키워드

과제정보

We are extremely grateful to Dr. Chitrita DebRoy, Senior Research Associate & Deirector, Gastroenteric Disease Center, Wiley Lab, Penn State University, USA and to Dr. Bhushan Jayarao, Associate Professor, Cooperative Extension, Department of Veterinary Science, Penn State University (USA) for their guidance and help provided in preparing the manuscript. The Department of Microbiology and Biotechnology, M.S. University, Vadodara, India, facilitated the electron microscopic studies. We are thankful to the National Dairy Development Board, Anand, Gujarat for providing the facilities used in the present work.

참고문헌

  1. Altmann K, Pyliotis NA, Mukkur TKS. A new method for the extraction and purification of K99 pili from enterotoxigenic Escherichia coli and their characterization. Biochem J 1982, 201, 505-513 https://doi.org/10.1042/bj2010505
  2. Batra HV, Chand P, Thillaikoothan P, Talwar GP. Coagglutination test with coloured Staphylococcus aureus for detection of brucella antigens in cattle brucellosis. Vet Rec 1987, 121, 65-66 https://doi.org/10.1136/vr.121.3.65
  3. Boedeker EC. Vaccines for enterotoxigenic Escherichia coli: current status. Curr Opin Gastroenterol 2005, 21, 15-19
  4. Chakraborty S, Deokule JS, Garg P, Bhattacharya SK, Nandy RK, Nair GB, Yamasaki S, Takeda Y, Ramamurthy T. Concomitant infection of enterotoxigenic Escherichia coli in an outbreak of cholera caused by Vibrio cholerae O1 and O139 in Ahmedabad, India. J Clin Microbiol 2001, 39, 3241-3246 https://doi.org/10.1128/JCM.39.9.3241-3246.2001
  5. de Graaf FK, Klemm P, Gaastra W. Purification, characterization, and partial covalent structure of Escherichia coli adhesive antigen K99. Infect Immun 1981, 33, 877-883
  6. Guinee PAM, Veldkamp J, Jansen WH. Improved minca medium for the detection of K99 antigen in calf enterotoxigenic strains of Escherichia coli. Infect Immun 1977, 15, 676-678
  7. Holley DL, Allen SD, Barnett BB. Enzyme-linked immunosorbent assay, using monoclonal antibody, to detect enterotoxic Escherichia coli K99 antigen in feces of dairy calves. Am J Vet Res 1984, 45, 2613-261
  8. Jay CM, Bhaskaran S, Rathore KS, Waghela SD. Enterotoxigenic K99+ Escherichia coli attachment to host cell receptors inhibited by recombinant pili protein. Vet Microbiol 2004, 101, 153-160 https://doi.org/10.1016/j.vetmic.2004.03.019
  9. Jiang ZD, Lowe B, Verenkar MP, Ashley D, Steffen R, Tornieporth N, von Sonnenburg F, Waiyaki P, DuPont HL. Prevalence of enteric pathogens among international travelers with diarrhea acquired in Kenya (Mombasa), India (Goa), or Jamaica (Montego Bay). J Infect Dis 2002, 185, 497-502 https://doi.org/10.1086/338834
  10. Kang G, Ramakrishna BS, Daniel J, Mathan M, Mathan VI. Epidemiological and laboratory investigations of outbreaks of diarrhoea in rural South India: implications for control of disease. Epidemiol Infect 2001, 127, 107-112
  11. Korhonen TK, Nurmiaho EL, Ranta H, Eden CS. New method for isolation of immunologically pure pili from Escherichia coli. Infect Immun 1980, 27, 569-575
  12. Kumar A, Sharma VD, Thapliyal DC. Occurrence of enterotoxigenic Escherichia coli in cow calves and infants/ children. Indian J Comp Microbiol Immunol Infect Dis 1982, 3, 174-177
  13. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970, 227, 680-685 https://doi.org/10.1038/227680a0
  14. Lugtenberg B, Meijers J, Peters R, van der Hoek P, van Alphen L. Electrophoretic resolution of the 'major outer membrane protein' of Escherichia coli K12 into four bands. FEBS Lett 1975, 58, 254-258 https://doi.org/10.1016/0014-5793(75)80272-9
  15. Micklem LR, Mc Cann MC, James K. The use of rat mixed-thymocyte culture-conditioned medium for hybridoma production, cloning and revival. J Immunol Methods 1987, 104, 81-86 https://doi.org/10.1016/0022-1759(87)90490-X
  16. Mills KW, Phillips RM, Kelly BL, Baughman GL. Using enzyme-linked immunosorbent assay to detect Escherichia coli K88 pili antigens from clinical isolates. Am J Vet Res 1982, 43, 365-367
  17. Morris JA, Stevens AE, Sojka WJ. Preliminary characterization of cell-free K99 antigen isolated from Escherichia coli B41. J Gen Microbiol 1977, 99, 353-357
  18. Raybould TJG, Crouch CF, Acres SD. Monoclonal antibody passive hemagglutination and capture enzymelinked immunosorbent assays for direct detection and quantitation of F41 and K99 fimbrial antigens in enterotoxigenic Escherichia coli. J Clin Microbiol 1987, 25, 278-284
  19. Rudin A, McConnell MM, Svennerholm AM. Monoclonal antibodies against enterotoxigenic Escherichia coli colonization factor antigen I (CFA/I) that cross-react immunologically with heterologous CFAs. Infect Immun 1994, 62, 4339-4346
  20. Sambrook J, Fritsch EF, Maniatis T. Molecular Cloning: A Laboratory Manual. 2nd ed. pp. 18.47-18.67, Cold Spring Harbor Laboratory Press, New York, 1989
  21. Sussman M. Escherichia coli in human and animal disease. In: Sussman M (ed.). The Virulence of Escherichia Coli: Review and Methods. pp. 7-45, Academic Press, London, 1985
  22. Tacket CO, Maneval DR, Levine MM. Purification, morphology, and genetics of a new fimbrial putative colonization factor of enterotoxigenic Escherichia coli O159:H4. Infect Immun 1987, 55, 1063-1069
  23. Thorns CJ, Sawtell JA, Miller JA, Wood GW. Identification of the K99 fimbrial adhesin in commercial vaccines used against calf enteritis. Biologicals 1990, 18, 113-115 https://doi.org/10.1016/1045-1056(90)90020-Z
  24. Tsen HY, Jian LZ. Development and use of a multiplex PCR system for the rapid screening of heat labile toxin I, heat stable toxin II and shiga-like toxin I and II genes of Escherichia coli in water. J Appl Microbiol 1998, 84, 585- 592 https://doi.org/10.1046/j.1365-2672.1998.00385.x
  25. Vazquez F, Gonzalez EA, Garabal JI, Blanco J. Fimbriae extracts from enterotoxigenic Escherichia coli strains of bovine and porcine origin with K99 and/or F41 antigens. Vet Microbiol 1996, 48, 231-241 https://doi.org/10.1016/0378-1135(95)00157-3
  26. Watterworth L, Topp E, Schraft H, Leung KT. Multiplex PCR-DNA probe assay for the detection of pathogenic Escherichia coli. J Microbiol Methods 2005, 60, 93-105 https://doi.org/10.1016/j.mimet.2004.08.016
  27. Woodward MJ, Wray C. Nine DNA probes for detection of toxin and adhesin genes in Escherichia coli isolated from diarrhoeal disease in animals. Vet Microbiol 1990, 25, 55-65 https://doi.org/10.1016/0378-1135(90)90093-B