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Serological Analysis by Enzyme-Linked Immunosorbent Assay Using Recombinant Antigen LipL32 for the Diagnosis of Swine Leptospirosis

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Abstract

Leptospirosis is an important global zoonotic disease caused by pathogenic Leptospira spp. species. Swine leptospirosis has a major economic impact because pigs are sources of animal protein and by-products. The signs of swine leptospirosis are abortion, stillbirth, birth of weak or ill piglets, appearing 14–60 days after infection. The reference method for diagnosis of leptospirosis is the microscopic agglutination test (MAT), in which serum samples are reacted with live antigen suspensions of leptospiral serovars. However, MAT is laborious and time consuming as a diagnostic procedure when dealing with a large number of samples; therefore, efforts are being made to develop novel, sensitive, and specific diagnostic tests for leptospirosis. In this study, a recombinant LipL32 based on enzyme-linked immunosorbent assay (rLipL32/ELISA) was evaluated as a screening test for the detection of pathogenic leptospiral-specific antibodies. A total of 86 swine serum samples tested by MAT were used to develop rLipL32/ELISA. Compared to positive and negative sera tested by MAT, rLipL32/ELISA showed 100 % sensitivity, 85.1 % specificity, and 91.86 % accuracy. No positive reaction for other bacterial diseases (enzootic pneumonia and brucellosis) was observed. The rLipL32/ELISA reported in this study is a specific, sensitive, and convenient test for the detection of antibodies against swine leptospiral infection and can be used as a rapid screening test in epidemiological surveys.

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References

  1. Adler B, de la Pena MA (2010) Leptospira and leptospirosis. Vet Microbiol 140:287–296

    Article  PubMed  CAS  Google Scholar 

  2. Adler B, Lo M, Seemann T et al (2011) Pathogenesis of leptospirosis: the influence of genomics. Vet Microbiol 153:73–81

    Article  PubMed  CAS  Google Scholar 

  3. Arimitsu Y, Kmety E, Ananyina Y et al (1994) Evaluation of the one-point microcapsule agglutination-test for diagnosis of leptospirosis. Bull World Health Organ 72:395–399

    PubMed  CAS  Google Scholar 

  4. Bomfim MR, Ko A, Koury MC (2005) Evaluation of the recombinant LipL32 in enzyme-linked immunosorbent assay for the serodiagnosis of bovine leptospirosis. Vet Microbiol 109:89–94

    Article  PubMed  CAS  Google Scholar 

  5. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  PubMed  CAS  Google Scholar 

  6. Brandao AP, Camargo ED, da Silva ED et al (1998) Macroscopic agglutination test for rapid diagnosis of human leptospirosis. J Clin Microbiol 36:3138–3142

    PubMed  CAS  Google Scholar 

  7. Cullen PA, Cordwell SJ, Bulach DM et al (2002) Global analysis of outer membrane proteins from Leptospira interrogans serovar Lai. Infect Immun 70:2311–2318

    Article  PubMed  CAS  Google Scholar 

  8. Cullen PA, Haake DA, Adler B (2004) Outer membrane proteins of pathogenic spirochetes. FEMS Microbiol Rev 28:291–318

    Article  PubMed  CAS  Google Scholar 

  9. Cumberland P, Everard CO, Levett PN (1999) Assessment of the efficacy of an IgM-elisa and microscopic agglutination test (MAT) in the diagnosis of acute leptospirosis. Am J Trop Med Hyg 61:731–734

    PubMed  CAS  Google Scholar 

  10. Dey S, Mohan CM, Kumar TM et al (2004) Recombinant LipL32 antigen-based single serum dilution ELISA for detection of canine leptospirosis. Vet Microbiol 103:99–106

    Article  PubMed  CAS  Google Scholar 

  11. Effler PV, Domen HY, Bragg SL et al (2000) Evaluation of the indirect hemagglutination assay for diagnosis of acute leptospirosis in Hawaii. J Clin Microbiol 38:1081–1084

    PubMed  CAS  Google Scholar 

  12. Faine S, Adler B, Bolin C, Perolat P (1999) Leptospira and leptospirosis. MedSci, Melbourne

    Google Scholar 

  13. Flannery B, Costa D, Carvalho FP et al (2001) Evaluation of recombinant Leptospira antigen-based enzyme-linked immunosorbent assays for the serodiagnosis of leptospirosis. J Clin Microbiol 39:3303–3310

    Article  PubMed  CAS  Google Scholar 

  14. Guerreiro H, Croda J, Flannery B et al (2001) Leptospiral proteins recognized during the humoral immune response to leptospirosis in humans. Infect Immun 69:4958–4968

    Article  PubMed  CAS  Google Scholar 

  15. Haake DA, Matsunaga J (2010) Leptospira: a spirochaete with a hybrid outer membrane. Mol Microbiol 77:805–814

    CAS  Google Scholar 

  16. Hoke DE, Egan S, Cullen PA et al (2008) LipL32 is an extracellular matrix-interacting protein of Leptospira spp. and Pseudoalteromonas tunicata. Infect Immun 76:2063–2069

    Article  PubMed  CAS  Google Scholar 

  17. Hull-Jackson C, Glass MB, Ari MD et al (2006) Evaluation of a commercial latex agglutination assay for serological diagnosis of leptospirosis. J Clin Microbiol 44:1853–1855

    Article  PubMed  CAS  Google Scholar 

  18. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Article  PubMed  CAS  Google Scholar 

  19. Levett PN (2001) Leptospirosis. Clin Microbiol Rev 14:296–326

    Article  PubMed  CAS  Google Scholar 

  20. Lin X, Zhao J, Qian J et al (2010) Identification of immunodominant B- and T-cell combined epitopes in outer membrane lipoproteins LipL32 and LipL21 of Leptospira interrogans. Clin Vaccin Immunol 17:778–783

    Article  CAS  Google Scholar 

  21. Mariya R, Chaudhary P, Kumar AA et al (2006) Evaluation of a recombinant LipL41 antigen of Leptospira interrogans serovar canicola in ELISA for serodiagnosis of bovine leptospirosis. Comp Immunol Microbiol Infect Dis 29:269–277

    Article  PubMed  CAS  Google Scholar 

  22. Merien F, Truccolo J, Baranton G et al (2000) Identification of a 36-kDa fibronectin-binding protein expressed by a virulent variant of Leptospira interrogans serovar icterohaemorrhagiae. FEMS Microbiol Lett 185:17–22

    Article  PubMed  CAS  Google Scholar 

  23. Rajeev S, Berghaus RD, Overton MW et al (2010) Comparison of fluorescent antibody and microscopic agglutination testing for Leptospira in pregnant and nonpregnant cows. J Vet Diagn Invest 22:51–54

    Article  PubMed  Google Scholar 

  24. Smits HL, Eapen CK, Sugathan S et al (2001) Lateral-flow assay for rapid serodiagnosis of human leptospirosis. Clin Diagn Lab Immunol 8:166–169

    PubMed  CAS  Google Scholar 

  25. Wang Z, Jin L, Wegrzyn A (2007) Leptospirosis vaccines. Microb Cell Fact 6:39

    Article  PubMed  Google Scholar 

  26. Wasinski B, Pejsak Z (2010) Occurrence of leptospiral infections in swine population in Poland evaluated by ELISA and microscopic agglutination test. Pol J Vet Sci 13:695–699

    PubMed  CAS  Google Scholar 

  27. Wolff JW (1954) The laboratory diagnosis of leptospirosis. Charles C Thomas, Springfield

    Google Scholar 

Download references

Acknowledgments

This study was supported by FAPERGS Programa Pesquisador Gaúcho—Edital PqG No. 06/2010, Grant 1010724.

Conflict of interest

No competing financial interests exist.

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Authors and Affiliations

  1. Laboratório de Imunodiagnóstico, Núcleo de Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, P.O. Box 354, Pelotas, RS, CEP 96010-900, Brazil

    Cláudia P. Hartleben, Fernanda M. A. Leal & Leonardo G. Monte

  2. Laboratório de Vacinologia, Núcleo de Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, P.O. Box 354, Pelotas, RS, CEP 96010-900, Brazil

    Daiane D. Hartwig & Odir A. Dellagostin

  3. Laboratório de Genômica Funcional, Núcleo de Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, P.O. Box 354, Pelotas, RS, CEP 96010-900, Brazil

    Fabiana K. Seixas

  4. Departamento de Medicina Veterinária Preventiva e Saúde Animal da Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo, São Paulo, SP, Brazil

    Sílvio A. Vasconcellos

  5. Instituto de Patobiologia, Centro de Ciencias Veterinarias, y Agronômicas (CICVyA), Instituto Nacional de Tecnologia Agropecuaria (INTA), Buenos Aires, Argentina

    Bibiana Brihuega

Authors
  1. Cláudia P. Hartleben
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  2. Fernanda M. A. Leal
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  3. Leonardo G. Monte
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  4. Daiane D. Hartwig
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  5. Fabiana K. Seixas
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  6. Sílvio A. Vasconcellos
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  7. Bibiana Brihuega
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  8. Odir A. Dellagostin
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Corresponding author

Correspondence to Cláudia P. Hartleben.

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Hartleben, C.P., Leal, F.M.A., Monte, L.G. et al. Serological Analysis by Enzyme-Linked Immunosorbent Assay Using Recombinant Antigen LipL32 for the Diagnosis of Swine Leptospirosis. Curr Microbiol 66, 106–109 (2013). https://doi.org/10.1007/s00284-012-0237-x

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  • DOI: https://doi.org/10.1007/s00284-012-0237-x

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