Evaluation of real-time PCR, MAT, and recombinant LipL32-based ELISA for the diagnosis of canine leptospirosis in a disease-endemic South Indian state, Kerala

Canine samples (n = 425) collected from cases of suspected leptospirosis over a period of 6 years were tested using the microscopic agglutination test (MAT), recombinant LipL32-based IgM and IgG ELISA, and real-time PCR (RT PCR). A prevalence of 69.41% was recorded by MAT. Among the different tests, IgG ELISA was observed to have maximum agreement with that of MAT (k-0.810), followed by IgM ELISA (k-0.762.). RT PCR had only slight agreement with MAT (k-0.156). However, the percentage positivity of RT PCR with samples with MAT titers of 1:100 and 1:200 were 76.47% and 85.96%, respectively, indicating the reliability of RT PCR in the early phase of the disease. The values for the samples at higher MAT titers of 1:400 and 1:800 were 10.58% and 1.2%, respectively, clearly indicating the unreliability of the test for samples having high antibody titers. It was concluded that IgG ELISA could be used as the routine test for the diagnosis of canine leptospirosis in this disease-endemic region. Samples with negative results that have high symptomatic correlation with leptospirosis may be tested by PCR/RT PCR for the detection of the pathogen well ahead of antibody generation.

___

  • Adler B, Moctezuma A. Leptospira and leptospirosis. Vet Microbiol 2010; 140: 287-296.
  • Branger C, Blanchard B, Fillonneau C, Suard I, Aviat F, Chevallier B, André‐Fontaine G. Polymerase chain reaction assay specific for pathogenic Leptospira based on the gene hap1 encoding the hemolysis‐associated protein‐1. FEMS Microbiol Lett 2005; 243: 437-445.
  • Joseph S, Thomas N, Thangapandian E, Singh VP, Verma R, Srivastava SK. Evaluation and comparison of native and recombinant LipL21 protein-based ELISAs for diagnosis of bovine leptospirosis. J Vet Sci 2012; 13: 99-101.
  • Bourhahy P, Bremont S, Zinini F, Giry C, Picardeau M. Comparison of real- time PCR assays for the detection of pathogenic Leptospira spp. in blood and identification of variations in target sequences. J Clin Microbiol 2011; 49: 2154- 2160.
  • Waggoner JJ, Balassiano I, Abeynayke J, Sahoo MK, Mohamed- Hadley A, Liu Y, Vital-Brazil MJ, Pinsky AB. Sensitive real-time PCR detection of pathogenic Leptospira spp and a comparison of nucleic acid amplification methods for the diagnosis of leptospirosis. PLoS One 2014; 9: e 112356.
  • Denipitiya DTH, Chandrasekharan NV, Abeyewickreme W, Hartkeerl CM, Hartskeerl RA, Jiffrey AM, Hapugoda MD. Application of a real time polymerase chain reaction (PCR) assay for the early diagnosis of human leptospirosis in Sri Lanka. Biologicals 2016; 44: 497-502.
  • Dey S, Mohan CM, Kumar TM, Ramadass P, Nainar AM, Nachimuthu K. Recombinant LipL32 antigen-based single serum dilution ELISA for detection of canine leptospirosis. Vet Microbiol 2004; 103: 99-106.
  • Deneke Y, Sabarinath T, Gogia N, Lasiamthara J, Viswas KN, Chaudhuri P. Evaluation of recombinant LigB antigen- based indirect ELISA and latex agglutination test for the serodiagnosis of bovine leptospirosis in India. Mol Cell Probe 2014; 28: 141-146.
  • Budhihal VS, Perwez K. Leptospirosis diagnosis: competancy of various laboratory tests. J Clin Diagnostic Res 2014; 8: 199- 202.
  • Guerreiro H, Croda J, Flannery B, Mazel M, Matsunaga J, Reis MG, Levett PN, Ko AI, Haake DA. Leptospiral proteins recognized during the humoral immune response to leptospirosis in human. Infect Immun 2001; 69: 4958-4968.
  • Ooteman MC, Vago AR, Koury MC. Evaluation of MAT, IgM ELISA and PCR methods for diagnosis of human leptospirosis. J Microbiol 2006; 65: 247-257.
  • Goris MG, Hartskeerl RA. Leptospirosis serodiagnosis by the microscopic agglutination test. Curr Protoc Microbiol 2014; 6: 32-35.
  • Rajapakse S, Weeratunga P, Niloofa R, Fernando N, de Silva LN, Rodrigo C, Maduranga S, Nandasir N, Premawansa S, Karunanayake L et al. A diagnostic scoring model for leptospirosis in resource limited settings. PLoS Negl Trop Dis 2016; 10: e0004513.
  • Niloofa R, Fernando N, de Silva NL, Karunanayake L, Wickramasinghe H, Dikmadugoda N, Premawansa G, Wickramasinghe R, de Silva JH, Premawansa S et al. Diagnosis of leptospirosis: comparison between microscopic agglutination test, IgM-ELISA and IgM rapid immunochromatography test. PLoS One 2015; 10: e0129236.
Turkish Journal of Veterinary and Animal Sciences-Cover
  • ISSN: 1300-0128
  • Yayın Aralığı: Yılda 6 Sayı
  • Yayıncı: TÜBİTAK