Evaluation of the diagnostic potential and DIVA capability of recombinant LigBCon1-5 protein of Leptospira interrogans serovar Pomona in canine leptospirosis

Document Type : Full paper (Original article)

Authors

1 Department of Epidemiology & Public Health, School of Life Science, Central University of Tamil Nadu, Tamil Nadu, India

2 Clinical Bacteriological Laboratory, ICAR-Indian Veterinary Research Institute (ICAR-IVRI), Mukteshwar, India

3 Department of Microbial Biotechnology, School of Veterinary Medicine, Jimma University, Jimma, Ethiopia

4 Department of Veterinary Medicine, Veterinary Clinical Complex, G C Negi College of Veterinary & Animal Sciences, Chaudhary Sarwan Kumar HP Krishi University, Palampur, Himachal Pradesh, India

5 Veterinary Medicine Division, ICAR-Indian Veterinary Research Institute (ICAR-IVRI), Izatnagar, Bareilly, India

6 Division of Veterinary Public Health, ICAR-Indian Veterinary Research Institute (ICAR-IVRI), Izatnagar, Bareilly, India

7 Zoonosis Research Lab, Madras Veterinary College, Tamil Nadu Veterinary and Animal Sciences University, Tamil Nadu, India

8 Livestock Economics & Statistics Division, ICAR-Indian Veterinary Research Institute (ICAR-IVRI), Izatnagar, Bareilly, India

9 Biochemistry Laboratory, ICAR-Indian Veterinary Research Institute (ICAR-IVRI), Mukteswar, India

10 Ph.D. Student in Biotechnology, Institute of Chemistry, Academia Sinica University, Nankang, Taipei, Taiwan

Abstract

Background: Canine leptospirosis is a serious public health concern. Aims: This study aims to investigate the feasibility of conserved first to fifth domains of recombinant Leptospira immunoglobulin like protein B antigen (rLigBCon1-5) as a serodiagnostic marker for detecting canine leptospirosis. Methods: A total of 340 unvaccinated canine serum samples were screened using microscopic agglutination test (MAT) and rLigBCon1-5 based immunoglobulin G (IgG) indirect-enzyme-linked immunosorbent assay (I-ELISA). Further, 60 vaccinated canine sera were screened using MAT and rLigBCon1-5 based latex agglutination test (LAT). Results: Microscopic agglutination test results revealed seropositivity of 28.6%. The relative sensitivity, specificity, and accuracy of IgG I-ELISA in comparison to MAT were 100%, 96.0%, and 97.2%, respectively. Out of 60 vaccinated sera, 46 sera reacted with MAT alone, and eight sera reacted by both tests, while six sera were non-reactive with both tests. Anti-LigB antibodies were detected in eight canine sera by rLigBCon1-5 based LAT. In five LAT reactive sera, agglutinins of locally circulating Leptospira serovars Grippotyphosa (n=4) and Australis (n=1) were detected. In three LAT reactive sera, agglutinins against Icterohaemorrhagiae (n=3) produced due to natural infection were present. Conclusion: Immunoglobulin G based indirect ELISA assay (IgG I-ELISA) can be employed as an alternative test instead of MAT. rLigBCon1-5 based LAT detected anti-LigB antibodies in eight vaccinated sera where the vaccine failure occurred partially or totally due to the limited efficacy spectrum of Nobivac® RL and cold chain breakage. This vaccine could not provide cross-protection against locally circulating Leptospira serovars. The recombinant LigBCon1-5 antigen based LAT possesses capability of differentiating infected from vaccinated individuals (DIVA capability) when employed as a pen-side test for detecting canine leptospirosis.

Keywords

Main Subjects

References

Abdollahpour, G; Shafighi, T and Sattari Tabrizi, S (2009). Serodiagnosis of leptospirosis in cattle in North of Iran, Gilan. Int. J. Vet. Res., 3: 7-10.
Adler, B (2015). Vaccines against leptospirosis: Leptospira and leptospirosis. Curr. Top. Microbiol. Immunol., 387: 251-272.
Alexander, AD (1986). Serological diagnosis of leptospirosis. In: Rose, NR; Friedman, H and Fahey, JL (Eds.), Manual of clinical laboratory immunology. (3rd Edn.), American Society for Microbiology, Washington D. C., PP: 435-439.
Al-orry, W; Arahou, M; Hassikou, R; Quasmaoui, A; Charof, R and Mennane, Z (2016). Leptospirosis: transmission, diagnosis and orevention. IJIAS. 15: 457-467.
Ambily, R; Mini, M; Joseph, S; Krishna, SV and Abhinay, G (2013). Canine leptospirosis – a seroprevalence study from Kerala, India. Vet. World. 6: 42-44.
André-Fontaine, G (2016). Leptospirosis in domestic animals in France: serological results from 1988 to 2007. Rev. Sci. Tech., 35: 913-923.
Budihal, SV and Perwez, K (2014). Leptospirosis diagnosis: competancy of various laboratory tests. J. Clin. Diagn. Res., 8: 199-202.
Claus, A; Van de maele, I; Gommeren, K; Pasmans, F and Daminet, S (2008). Leptospirosis in dogs: a retrospective study of seven clinical cases in Belgium. Vlaams Diergeneeskd. Tijdschr., 77: 259-263.
Croda, J; Ramos, JG; Matsunaga, J; Queiroz, A; Homma, A; Riley, LW; Haake, DA; Reis, MG and Ko, AI (2007). Leptospira immunoglobulin-like proteins as a serodiagnostic marker for acute leptospirosis. J. Clin. Microbiol., 45: 1528-1534.
Day, MJ; Horzinek, MC; Schultz, RD and Squires, RA (2016). WSAVA guidelines for the vaccination of dogs and cats. J. Small Anim. Pract., 57: E1-E45.
Deneke, Y; Sabarinath, T; Gogia, N; Lalsiamthara, J; Viswas, KN and Chaudhuri, P (2014). Evaluation of recombinant LigB antigen-based indirect ELISA and latex agglutination test for the serodiagnosis of bovine leptospirosis in India. Mol. Cell. Probes. 28: 141-146.
de Savigny, D and Voller, A (1980). The communication of ELISA data from laboratory to clinician. J. Immunoassay. 1: 105-128.
Doosti, A; Ahmadi, R and Arshi, A (2012). PCR detection of leptospirosis in Iranian Camels. Bulg. J. Vet. Med., 15: 178-183.
Engvall, E and Perlmann, P (1971). Enzyme-linked immunosorbent assay (ELISA). Quantitative assay of immunoglobulin G. Immunochemistry. 8: 871-874.
Feigin, RD; Lobes, LA; Anderson, D and Pickering, L (1973). Human leptospirosis from immunized dogs. Ann. Intern. Med., 79: 777-785.
Friedrich, K and Truyen, U (2000). Efficacy of parvovirus vaccines and effectiveness of two vaccination protocols. Prakt. Tierarzt., 81: 988-994.
Greene, CE; Sykes, JE; Brown, CA and Hartmann, K (2006). Leptospirosis. In: Greene, CE (Ed.), Infectious diseases of the dog and cat. (3rd Edn.), W. B. Saunders, St Louis. PP: 402-417.
Hartleben, CP; Leal, FM; Monte, LG; Hartwig, DD; Seixas, FK; Vasconcellos, SA; Brihuega, B and Dellagostin, OA (2013). Serological analysis by enzyme-linked immunosorbent assay using recombinant antigen LipL32 for the diagnosis of swine leptospirosis. Curr. Microbiol., 66: 106-109.
Higino, SSS; Azevedo, SS; Alves, CJ; Figueiredo, SM; Silva, MLCR and Batista, CSA (2010). Frequency of leptospirosis in sheep slaughtered in the municipality of Patos, Paraíba. Arq. Inst. Biol., 77: 525-527.
Jacobson, RH (1998). Validation of serological assays for diagnosis of infectious diseases. Rev. Sci. Tech. Off. Int. Epiz., 17: 469-486.
Klaasen, HE and Adler, B (2015). Recent advances in canine leptospirosis: focus on vaccine development. Vet. Med. (Auckl)., 6: 245-260.
Laemmli, UK (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 227: 680-685.
Lau, CL; Skelly, C; Smythe, LD; Craig, SB and Weinstein, P (2012). Emergence of new leptospiral serovars in American Samoa-ascertainment or ecological change? BMC Infect. Dis., 12: 19-26.
Levett, PN (2001). Leptospirosis. Clin. Microbiol. Rev., 14: 296-326.
Lin, X; Chen, Y and Yan, J (2008). Recombinant multiepitope protein for diagnosis of leptospirosis. Clin. Vaccine Immunol., 15: 1711-1714.
Matsunaga, J; Barocchi, MA; Croda, J; Young, TA; Sanchez, Y; Siqueira, I; Bolin, CA; Reis, MG; Riley, LW; Haake, DA and Ko, AI (2003). Pathogenic Leptospira species express surface-exposed proteins belonging to the bacterial immunoglobulin superfamily. Mol. Microbiol., 49: 929-945.
Musso, D and Scola, BL (2013). Laboratory diagnosis of leptospirosis: A challenge. J. Microbiol. Immunol. Infect., 46: 245-252.
Palaniappan, RU; Chang, YF; Hassan, F; McDonough, SP; Pough, M; Barr, SC; Simpson, KW; Mohammed, HO; Shin, S; McDonough, P; Zuerner, RL; Qu, J and Roe, B (2004). Expression of leptospiral immunoglobulin-like protein by Leptospira interrogans and evaluation of its diagnostic potential in a kinetic ELISA. J. Med. Microbiol., 53: 975-984.
Patil, D; Dahake, R; Roy, S; Mukherjee, S; Chowdhary, A and Deshmukh, R (2014). Prevalence of leptospirosis among dogs and rodents and their possible role in human lepto spirosis from Mumbai, India. Indian J. Med. Microbiol., 32: 64-67.
Sabarinath, T; Behera, SK; Deneke, Y; Atif Ali, S; Kaur, G; Ashok Kumar, T; Ravi Kumar, G; Senthil Kumar, K; Sinha, DK; Verma, MR; Srivastava, SK and Chaudhuri, P (2018). Serological evidence of anti-Leptospira antibodies in goats in various agro climatic zones of India. Small Rumin. Res., 169: 74-80.
Sathiyamoorthy, A; Selvaraju, G; Palanivel, KM and Srinivasan, P (2017). Seroprevalence of canine leptospirosis in Namakkal, Tamil Nadu by Microscopic Agglutination Test. J. Cell Tissue Res., 17: 5991-5996.
Senthilkumar, T; Subathra, M; Phil, M; Ramadass, P and Ramaswamy, V (2008). Rapid serodiagnosis of leptospirosis by latex agglutination test and flow-through assay. Indian J. Med. Microbiol., 26: 45-49.
Smits, HL; van der Hoorn, MA; Goris, MG; Gussenhoven, GC; Yersin, C and Sasaki, DM (2000). Simple latex agglutination assay for rapid serodiagnosis of human leptospirosis. J. Clin. Microbiol., 38: 1272-1275.
Srikram, A; Zhang, K; Bartpho, T; Lo, M; Hoke, DE; Sermswan, RW; Adler, B and Murray, GL (2011). Cross-protective immunity against leptospirosis elicited by a live, attenuated lipopolysaccharide mutant. J. Infect. Dis., 203: 870-879.
Stokes, JE and Forrester, SD (2004). New and unusual causes of acute renal failure in dogs and cats. Vet. Clin. North Am. Small Anim. Pract., 34: 909-922.
Towbin, H; Staehelin, T and Gordon, J (1979). Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc. Natl. Acad. Sci. USA., 76: 4350-4354.
van de Maele, I; Claus, A; Haesebrouck, F and Daminet, S (2008). Leptospirosis in dogs: a review with emphasis on clinical aspects. Vet. Rec., 163: 409-413.
Xu, Y; Zheng, H; Zhang, Y; Wang, Y; Zhang, J; Li, Z; Cui, S; Xin, X; Ye, Q; Chang, YF and Wang, J (2017). Genomic analysis of a new serovar of Leptospira weilii serogroup Manhao. Front. Microbiol., 8: 149.
Ye, C; Yan, W; Xiang, H; He, H; Yang, M; Ijaz, M; Useh, N; Hsieh, CL; McDonough, PL; McDonough, SP; Mohamed, H; Yang, Z and Chang, YF (2014). Recombinant antigens rLipL21, rLoa22, rLipL32 and rLigACon4-8 for serological diagnosis of leptospirosis by enzyme-linked immunosorbent assays in dogs. PLoS One. 9: e111367.

Files

Share

How to cite

Statistics