Preliminary Study: Characterisation of Antibody for rLipl32 Protein of Leptospira

Sumarningsih ., Simson Tarigan

Abstract


Leptospirosis infection is commonly determined by positive response antibody in microaglutination test (MAT). The test requires panels of Leptospira from complete serovars and uses infectious Leptospira, which is subcultured weekly. Accordingly, other alternative serological tests are developed for MAT. One of them is ELISA using recombinant LipL32 protein (rLipL32). High sensitivity and specificity of ELISA rLipL32 for Leptospirosis detection in bovine and canine has been reported previously. In this study, we produced antibody to rLipL32 protein with high titre. Results on western blotting and ELISA showed anti-LipL32 antibody was highly binding to rLipL32 protein. In contrast, MAT results for LipL32 antibody was found to be negative response, indicating that titre antibody to LipL32 does not positively correlate to MAT.

Keywords


Leptospira; Antibody Response; LipL32; MAT

References


Bajani MD, Ashford DA, Bragg SL, Woods CW, Aye T, Spiegel RA, Plikaytis BD, Perkins BA, Phelan M, Levett PN, Weyant RS. 2003. evaluation of four commercially available rapid serologic tests for diagnosis of leptospirosis. J Clin Microbiol. 41:803-809.

Bomfim MRQ, 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.

Chaemchuen S, Rungpragayphan S, Poovorawan Y, Patarakul K. 2011. Identification of candidate host proteins that interact with LipL32, the major outer membrane protein of pathogenic Leptospira, by random phage display peptide library. Vet Microbiol. 153:178-185.

Cousins DV, Robertson GM, Hustas L. 1985. The use of the enzyme-linked immunosorbent assay (ELISA) to detect the IgM and IgG antibody response to Leptospira interrogans serovars hardjo, pomona and tarassovi in cattle. Vet Microbiol. 10: 439-450.

Cullen PA, Xu X, Matsunaga J, Sanchez Y, Ko AI, Haake DA, Adler B. 2005. Surfaceome of Leptospira spp. Infect Immun. 73: 4853-4863.

Dey S, Mohan CM, Kumar TMAS, Ramadass P, Nainar AM, Nachimuthu K. 2004. Recombinant LipL32 antigen-based single serum dilution ELISA for detection of canine leptospirosis. Vet Microbiol. 103:99-106.

Flannery B, Costa D, Carvalho FP, Guerreiro H, Matsunaga J, Da Silva ED, Ferreira AGP, Riley LW, Reis MG, Haake DA, Ko AI. 2001. Evaluation of recombinant Leptospira antigen-based enzyme-linked immunosorbent assays for the serodiagnosis of leptospirosis. J Clin Microbiol. 39:3303-3310.

Haake DA, Chao G, Zuerner RL, Barnett JK, Barnett D, Mazel M, Matsunaga J, Levett PN, Bolin CA. 2000. The leptospiral major outer membrane protein LipL32 is a lipoprotein expressed during mammalian infection. Infect Immun. 68:2276-2285.

Levett PN. 2001. Leptospirosis. Clin Microbiol Rev. 14:296-326.

Malmström J, Beck M, Schmidt A, Lange V, Deutsch EW, Aebersold R. 2009. Proteome-wide cellular protein concentrations of the human pathogen Leptospira interrogans. Nature. 460:762-765.

Pinne M, Haake DA. 2013. LipL32 is a subsurface lipoprotein of Leptospira interrogans: Presentation of new data and reevaluation of previous studies. PLoS ONE. 8:e51025.

Seenichamy A, Bahaman AR, Mutalib AR, Khairani-Bejo S. 2014. Production and characterization of a polyclonal antibody of anti-rLipL21-IgG against Leptospira for early detection of acute leptospirosis. BioMed Research International. 2014:592858.

Sumarningsih, Tarigan S, Kusmiyati. 2016. Recombinant LipL32 protein for leptospirosis detection in Indonesia. Procedia Chemistry. 18:18-25.

OIE. 2013. Manual of diagnostic tests and vaccines for terrestrial animals. OIE. 4 Februari 2016, from http://www.oie.int/fileadmin/ Home/eng/Health_standards/tahm/2.01.09_LEP TO.pdf.

Yang CW, Wu MS, Pan MJ, Hsieh WJ, Vandewalle, Huang CC. 2002. The Leptospira outer membrane protein LipL32 induces tubulointerstitial nephritis-mediated gene expression in mouse proximal tubule cells. J Am Soc Nephrol. 13:2037-2045.




DOI: 10.14334/Proc.Intsem.LPVT-2016-p.147-152

Refbacks

  • There are currently no refbacks.