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Current IssueEditorial BoardOnline First ArticlesArchive

Research Article

volume 53 issue 1 (january 2019) : 99-104,   Doi: 10.18805/ijar.B-3382

Standardization and application of rapid tests viz dot ELISA and vertical flow through tests for serodiagnosis of classical swine fever 

A
Aparajita Das
D
Dilip K. Bhattacharyya
1Department of Veterinary Microbiology, College of Veterinary Sciences and Animal Husbandary, Assam Agriculture University, Khanapara, Guwahati-781 022, Assam, India.
Cite article:- Das Aparajita, Bhattacharyya K. Dilip (2018). Standardization and application of rapid tests viz dot ELISA and vertical flow through tests for serodiagnosis of classical swine fever. Indian Journal of Animal Research. 53(1): 99-104. doi: 10.18805/ijar.B-3382.

ABSTRACT

The present study was carried out to develop rapid field-based immunoassays for onsite detection of Classical swine fever virus (CSFV) antibody in clinical serum sample. Purified CSFV antigen was coated onto nitrocellulose membranes housed in a plastic module with layers of absorbent filter pads underneath. Following addition of serum to be tested appearance of a red dot indicated the presence of antibodies to CSF virus in the sample tested. A total of 240 serum samples collected from different areas of North Eastern States of India were tested for the presence of CSFV antibody by Indirect ELISA, Dot ELISA and Vertical flow through test. The result of the Indirect ELISA could be obtained within 2.5 hours whereas the result in case of Dot ELISA and Vertical Flow through test was obtained within 1 hour and 15 minutes respectively. Dot-ELISA and Vertical flow through test could equally detect the CSFV antibody in samples possessing 1:4 and above ELISA titre. The specificity and sensitivity of Dot-ELISA and vertical flow through test after comparison with Indirect ELISA indicated 100% specific and 84.28% sensitive. These tests have the potential to be used as a field-based kit to assess the serodiagnosis of CSFV.

REFERENCES

  1. Anonymous. (2002). European Union Diagnostic Manual on swinefever. Hanover. Germany.
  2. Black, D.N. and Brown, F. (1976) Purification and physicochemical characteristics of African swine fever virus. J Gen Virol., 32:509-    518.
  3. Clavijo, A., Lin, M., Riva, J. and Zhou, E. M. (2001). Application of competitive enzyme linked immunosorbent assay for the serologic diagnosis of classical swine fever virus infection. J. Vet. Diagn. Invest., 13: 357–360.
  4. Cui, S., Zhou, S., Chen, C., Qi, T., Zhang, C. and Oh, J. (2008). A simple and rapid immunochromatographic strip test for detecting antibody to porcine reproductive and respiratory syndrome virus. J. Virol. Met., 152: 38–42.
  5. Dhinakar, R.G., Thiagarajan, A.V. and Nachimuthu, K. (2007). Detection of antibodies to egg drop syndrome virus in chicken serum using a field-based immunofiltration (flow-through) test. Avian. Pathol., 51: 788–790.
  6. Farhid, H. and Farhad, A. (2009). Dot-blot enzyme immunoassay for the detection of bovine viral diarrhea virus antibodies. Veterinarski Arhiv., 79(4): 343-350.
  7. Handel, K., Kehler, H., Hills, K. and Pasick, J. (2004). Comparison of reverse transcriptase-polymerase chain reaction, virus isolation and immunoperoxidase assays for detecting pigs infected with low, moderate, and high virulent strains of classical swine fever virus. Journal of Veterinary Diagnostic Investigation., 16/2: 132-138.
  8. Hemmatzadeh, F.F. and Aminimmatzadeh, F.F. (2009). Dot-blot enzyme immunoassay for the detection Dot-blot enzyme immunoassay for the detection of bovine viral diarrhea virus antibodies. Vet. Arhiv., 79: 343-350.
  9. Karber, G. (1931). Beitrong Zur kollektiven Behandlung pharmakologischer Reihenversuche. Arch. Exp. Pathol. Pharmakol., 162: 480-483.
  10. Kripa, L.J., Niranjana, C., Kirubaharan, J.J., Danial, N. and Chandran, J. (2009). Development and standardization of an immunostick ELISA for rapid identification of Newcastle disease antibodies in chicken. Indian Vet. J., 86: 991-995.
  11. Leifer, I., Hoffmann, B., Hoper, D., Rasmuseen, T.B., Blome, S., Strebelow, G., Horeth-Bontgen, D., Staubach, C. and Beer, M. (2010). Molecular epidemiology of current classical swine fever virus isolates of wild boar in Germany. J. Gen. Virol .,91: 2687-2697.
  12. Malswamkima, D., Rajkhowa, T.K., Chandra, R. and Dutta, T.K. (2015). Pathology and molecular diagnosis of classical swine fever in Mizoram. Veterinary World., 8(1): 76-81.
  13. Natanael, L.D., Antônio, A.F., Anapolino, M.O., Érica, B.S., Bruna, R.C.A., Fernanda, A.D. and Marcelo, F.C. (2014). Validation of a Real Time PCR for classical swine fever Diagnosis. Veterinary Medicine International., 17: 12-35.
  14. OIE, (2014). Classical swine fever(Hog cholera)Infection with classical swine fever virus.In: OIE terrestrial manual, (Chapter 2.8.3).
  15. Rajbongshi, G., Barman, N. N., and Das, S. K. (2011). Survival and inactivation of classical swine fever virus isolated from pigs of Asom, India. Indian Journal of Animal Sciences., 81:330.
  16. Ross, R.N., Folds, D.J., Clifford, H. and Nakamura, M.R. (1997). Manual of Clinical Immunology., 5th Edn. p.24.
  17. Sarma, D.K. and Meshram, D.J. (2008). Comparison of Sandwich ELISA and Dot ELISA for detection of CSF virus antigen in pigs. Indian Vet J., 85: 915-918.
  18. Sarma, P.C. and Sarma, D.K. (1996). Use of indirect ELISA for detecting HCV antibodies in vaccinated piglets. Indian J. Anim. Sci., 66(11): 1128-1129.
  19. Sheng, D., Yuping, L. and Hong, Z. (2006). Dot-ELISA for detecting antibodies against classical swine fever in large scale pig farms. Guangxi Animal Husbandry and Veterinary Medicine.
  20. Shrivastva, A., Tripathi, N.K, Parida, M., Dash, P.K., Jana, A.M. and Lakshmana, R.P.V. (2008). Comparison of a dipstick enzyme-    linked immunosorbent assay with commercial assays for detection of Japanese encephalitis virus-specific IgM antibodies: Journal of Postgraduate Medicine., 22:181-185.
  21. Snedecor, G.W. and Cochran, W.G. (1994). Statistical Methods, 1st East-West Press ed., Affiliated East-West Private Ltd., New Delhi.
  22. Suzhen, Y., Jifei, Y., Gaiping, Z., Xuannian, W., Songlin, Q., Dong, Z., Yubao, Z., Xuewu, L., et al., (2010). Development of an immunochromatographic strip for the detection of antibodies against foot-and-mouth disease virus serotype O. Journal of Virological Methods.,165:139–144.
  23. Svobodova, Z., Jankovicova, B., Horak, D. and Bilkova, Z. (2013). Dot-ELISA Affinity Test: An easy, low-cost method to estimate binding activity of monoclonal antibodies. J Anal Bioanal Tech., 4:3.
  24. Talwar, G. P. (1983). A Handbook of Practical Immunology. 1st Edn., Vikas Publishing House Pvt. Ltd. pp. 71-72.
  25. Wang, B., Chen, J., Wang, M., Tian, F. and Jiang, S. (2010). The decreasing and increasing variance of CSF maternal antibodies in piglets. Chin. J. Vet. Sci., 30: 453–455.
  26. Xiu, J., Wu, S., Zhou, L., Ye, Y., Yu, D., Chen, R., Wang, L. and Zhang, X. (2010). Eradication and immunity evaluation on classic swine fever for pig farms of different sizes. Fujian J. Agric. Sci., 25: 235–238.
  27. Xuewu, L., Li, W., Xibao, S., Dong, Z., Jifei, S.Y. and Gaiping, Z. (2012). Development of an immunochromatographic strip for rapid detection of antibodies against classical swine fever virus. Journal of Virological Methods., 180: 32– 37.
  28. Yoshihiro, S., Hiroaki, W., Tehpin, T., Nomura, M.N., Hiroshi, H.M., Hiroshi, K. and Akio, F. (2012). Development and evaluation of indirect enzyme-linked immunosorbent assay for a screening test to detect antibodies against classical swine fever virus. Japanese Journal of Veterinary Research., 60(2&3): 85-94. 
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    Research Article

    volume 53 issue 1 (january 2019) : 99-104,   Doi: 10.18805/ijar.B-3382

    Standardization and application of rapid tests viz dot ELISA and vertical flow through tests for serodiagnosis of classical swine fever 

    A
    Aparajita Das
    D
    Dilip K. Bhattacharyya
    1Department of Veterinary Microbiology, College of Veterinary Sciences and Animal Husbandary, Assam Agriculture University, Khanapara, Guwahati-781 022, Assam, India.
    Cite article:- Das Aparajita, Bhattacharyya K. Dilip (2018). Standardization and application of rapid tests viz dot ELISA and vertical flow through tests for serodiagnosis of classical swine fever. Indian Journal of Animal Research. 53(1): 99-104. doi: 10.18805/ijar.B-3382.

    ABSTRACT

    The present study was carried out to develop rapid field-based immunoassays for onsite detection of Classical swine fever virus (CSFV) antibody in clinical serum sample. Purified CSFV antigen was coated onto nitrocellulose membranes housed in a plastic module with layers of absorbent filter pads underneath. Following addition of serum to be tested appearance of a red dot indicated the presence of antibodies to CSF virus in the sample tested. A total of 240 serum samples collected from different areas of North Eastern States of India were tested for the presence of CSFV antibody by Indirect ELISA, Dot ELISA and Vertical flow through test. The result of the Indirect ELISA could be obtained within 2.5 hours whereas the result in case of Dot ELISA and Vertical Flow through test was obtained within 1 hour and 15 minutes respectively. Dot-ELISA and Vertical flow through test could equally detect the CSFV antibody in samples possessing 1:4 and above ELISA titre. The specificity and sensitivity of Dot-ELISA and vertical flow through test after comparison with Indirect ELISA indicated 100% specific and 84.28% sensitive. These tests have the potential to be used as a field-based kit to assess the serodiagnosis of CSFV.

    REFERENCES

    1. Anonymous. (2002). European Union Diagnostic Manual on swinefever. Hanover. Germany.
    2. Black, D.N. and Brown, F. (1976) Purification and physicochemical characteristics of African swine fever virus. J Gen Virol., 32:509-    518.
    3. Clavijo, A., Lin, M., Riva, J. and Zhou, E. M. (2001). Application of competitive enzyme linked immunosorbent assay for the serologic diagnosis of classical swine fever virus infection. J. Vet. Diagn. Invest., 13: 357–360.
    4. Cui, S., Zhou, S., Chen, C., Qi, T., Zhang, C. and Oh, J. (2008). A simple and rapid immunochromatographic strip test for detecting antibody to porcine reproductive and respiratory syndrome virus. J. Virol. Met., 152: 38–42.
    5. Dhinakar, R.G., Thiagarajan, A.V. and Nachimuthu, K. (2007). Detection of antibodies to egg drop syndrome virus in chicken serum using a field-based immunofiltration (flow-through) test. Avian. Pathol., 51: 788–790.
    6. Farhid, H. and Farhad, A. (2009). Dot-blot enzyme immunoassay for the detection of bovine viral diarrhea virus antibodies. Veterinarski Arhiv., 79(4): 343-350.
    7. Handel, K., Kehler, H., Hills, K. and Pasick, J. (2004). Comparison of reverse transcriptase-polymerase chain reaction, virus isolation and immunoperoxidase assays for detecting pigs infected with low, moderate, and high virulent strains of classical swine fever virus. Journal of Veterinary Diagnostic Investigation., 16/2: 132-138.
    8. Hemmatzadeh, F.F. and Aminimmatzadeh, F.F. (2009). Dot-blot enzyme immunoassay for the detection Dot-blot enzyme immunoassay for the detection of bovine viral diarrhea virus antibodies. Vet. Arhiv., 79: 343-350.
    9. Karber, G. (1931). Beitrong Zur kollektiven Behandlung pharmakologischer Reihenversuche. Arch. Exp. Pathol. Pharmakol., 162: 480-483.
    10. Kripa, L.J., Niranjana, C., Kirubaharan, J.J., Danial, N. and Chandran, J. (2009). Development and standardization of an immunostick ELISA for rapid identification of Newcastle disease antibodies in chicken. Indian Vet. J., 86: 991-995.
    11. Leifer, I., Hoffmann, B., Hoper, D., Rasmuseen, T.B., Blome, S., Strebelow, G., Horeth-Bontgen, D., Staubach, C. and Beer, M. (2010). Molecular epidemiology of current classical swine fever virus isolates of wild boar in Germany. J. Gen. Virol .,91: 2687-2697.
    12. Malswamkima, D., Rajkhowa, T.K., Chandra, R. and Dutta, T.K. (2015). Pathology and molecular diagnosis of classical swine fever in Mizoram. Veterinary World., 8(1): 76-81.
    13. Natanael, L.D., Antônio, A.F., Anapolino, M.O., Érica, B.S., Bruna, R.C.A., Fernanda, A.D. and Marcelo, F.C. (2014). Validation of a Real Time PCR for classical swine fever Diagnosis. Veterinary Medicine International., 17: 12-35.
    14. OIE, (2014). Classical swine fever(Hog cholera)Infection with classical swine fever virus.In: OIE terrestrial manual, (Chapter 2.8.3).
    15. Rajbongshi, G., Barman, N. N., and Das, S. K. (2011). Survival and inactivation of classical swine fever virus isolated from pigs of Asom, India. Indian Journal of Animal Sciences., 81:330.
    16. Ross, R.N., Folds, D.J., Clifford, H. and Nakamura, M.R. (1997). Manual of Clinical Immunology., 5th Edn. p.24.
    17. Sarma, D.K. and Meshram, D.J. (2008). Comparison of Sandwich ELISA and Dot ELISA for detection of CSF virus antigen in pigs. Indian Vet J., 85: 915-918.
    18. Sarma, P.C. and Sarma, D.K. (1996). Use of indirect ELISA for detecting HCV antibodies in vaccinated piglets. Indian J. Anim. Sci., 66(11): 1128-1129.
    19. Sheng, D., Yuping, L. and Hong, Z. (2006). Dot-ELISA for detecting antibodies against classical swine fever in large scale pig farms. Guangxi Animal Husbandry and Veterinary Medicine.
    20. Shrivastva, A., Tripathi, N.K, Parida, M., Dash, P.K., Jana, A.M. and Lakshmana, R.P.V. (2008). Comparison of a dipstick enzyme-    linked immunosorbent assay with commercial assays for detection of Japanese encephalitis virus-specific IgM antibodies: Journal of Postgraduate Medicine., 22:181-185.
    21. Snedecor, G.W. and Cochran, W.G. (1994). Statistical Methods, 1st East-West Press ed., Affiliated East-West Private Ltd., New Delhi.
    22. Suzhen, Y., Jifei, Y., Gaiping, Z., Xuannian, W., Songlin, Q., Dong, Z., Yubao, Z., Xuewu, L., et al., (2010). Development of an immunochromatographic strip for the detection of antibodies against foot-and-mouth disease virus serotype O. Journal of Virological Methods.,165:139–144.
    23. Svobodova, Z., Jankovicova, B., Horak, D. and Bilkova, Z. (2013). Dot-ELISA Affinity Test: An easy, low-cost method to estimate binding activity of monoclonal antibodies. J Anal Bioanal Tech., 4:3.
    24. Talwar, G. P. (1983). A Handbook of Practical Immunology. 1st Edn., Vikas Publishing House Pvt. Ltd. pp. 71-72.
    25. Wang, B., Chen, J., Wang, M., Tian, F. and Jiang, S. (2010). The decreasing and increasing variance of CSF maternal antibodies in piglets. Chin. J. Vet. Sci., 30: 453–455.
    26. Xiu, J., Wu, S., Zhou, L., Ye, Y., Yu, D., Chen, R., Wang, L. and Zhang, X. (2010). Eradication and immunity evaluation on classic swine fever for pig farms of different sizes. Fujian J. Agric. Sci., 25: 235–238.
    27. Xuewu, L., Li, W., Xibao, S., Dong, Z., Jifei, S.Y. and Gaiping, Z. (2012). Development of an immunochromatographic strip for rapid detection of antibodies against classical swine fever virus. Journal of Virological Methods., 180: 32– 37.
    28. Yoshihiro, S., Hiroaki, W., Tehpin, T., Nomura, M.N., Hiroshi, H.M., Hiroshi, K. and Akio, F. (2012). Development and evaluation of indirect enzyme-linked immunosorbent assay for a screening test to detect antibodies against classical swine fever virus. Japanese Journal of Veterinary Research., 60(2&3): 85-94. 
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