Lateral flow assay for rapid serodiagnosis of bovine leptospirosis

Document Type : Full paper (Original article)

Authors

Zoonoses Research Laboratory, Centre for Animal Health Studies, Tamil Nadu Veterinary and Animal Sciences University, Chennai 600051, Tamil Nadu, India

Abstract

Background: Leptospirosis is considered to be an economically important disease in bovine. The disease burden is not appropriately monitored due to cumbersome serological tests that could be performed only in established laboratories. This warrants the development of a field level rapid diagnostic test. Aims: The study aimed to develop a lateral flow assay (LFA)-based pen-side diagnostic test to detect antibodies to Leptospira. Methods: LFA strip was prepared with the heat extracted antigen from L. interrogans serovar Pomona. To assess the performance of the developed LFA, a total of 300 bovine serum samples with their clinical histories were used and the initial screening for Leptospira antibodies was performed by the standard microscopic agglutination test (MAT). The sensitivity, specificity, and agreement (kappa value) were calculated between developed LFA and MAT. The stability of LFA was evaluated on days 30, 60, 90, and 120. Results: Out of 300 samples tested, 225 were positive, and 75 were negative on MAT and 208 were positive, and 92 were negative on LFA. The developed LFA had a sensitivity of 90.7% and a specificity of 94.7%. The results of the assay were substantially in agreement with MAT, with a kappa value of 0.79. The LFA strips were stable for 120 days at 4°C. Conclusion: A Lateral flow assay-based rapid pen-side test was developed and its utility to diagnose bovine leptospirosis was evaluated.

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Appassakij, H; Silpapojakul, K; Wansit, R and Woodtayakorn, J (1995). Evaluation of the immuno-fluorescent antibody test for the diagnosis of human leptospirosis. Am. J. Trop. Med. Hyg., 52: 340-343.
Behera, SK; Sabarinath, T; Deneke, Y; Bansal, SK; Mahendran, K; Kumar, A; Senthil, K; Verma, MR; ChandraSekar, S and Atif Ali, S (2021). Evaluation of the diagnostic potential and DIVA capability of recombinant LigBCon1-5 protein of Leptospira interrogans serovar Pomona in canine leptospirosis. Iran. J. Vet. Res., 22: 120-128.
Biswal, BC; Kar, BC; Ratnam, S; Sureshbabu, L and Natarajaseenivasan, K (2000). Evidence of antibodies to leptospira among farm cattle and farm workers in Orissa state. Ind. Vet. J., 77: 622-623.
Caimia, K and Ruybal, P (2020). Leptospira spp., a genus in the stage of diversity and genomic data expansion. Infect. Genet. Evol., https:/doi.org/10.1016/j.meegid.2020.104241.
Dikken, H and Kmety, E (1978). Serological typing methods of leptospires. Methods Microbiol., 11: 259-294.
Doungchawee, G; Sutdan, D; Niwatayakul, K; Inwisai, T; Sitthipunya, A; Boonsathorn, N; Sakulterdkiat, T; Sirawaraporn, W and Thongboongerd, V (2017). Development and evaluation of an immuno-chromatographic assay to detect serum anti-leptospiral lipopolysaccaride IgM in acute leptospirosis. Sci. Rep., 7: 2309. doi: 10.1038/s41598-017-02654-8.
Faine, S (1982). Guidelines for the control of Leptospirosis. World Health Organization, Geneva. P: 17.
Gussenhoven, GC; Van der hoorn, MAWG; Goris, MGA; Terpstra, WJ; Hartskeerl, RA; Mol, BW; Van ingen, CW and Smits, HL (1997). Lepto dipstick, a dipstick assay for detection of Leptospira-specific immunoglobulin M antibodies in human sera. J. Clin. Microbiol., 35: 92-97.
Jai sunder, T; Sujatha, A; Kundu, M and Kundu, S (2018). Carrier status and seroprevalence of leptospirosis in cattle of South Andaman. Ind. J. Anim. Res., 52: 140-143.
Levett, PN and Whittington, CU (1998). Evaluation of the indirect hemagglutination assay for diagnosis of acute leptopsirosis. J. Clin. Microbiol., 36: 11-14.
Maze, MJ; Sharples, KJ; Allan, KJ; Rubach, MP and Crump, JA (2019). Diagnostic accuracy of leptospirosis whole-cell lateral flow assays: a systemic review and meta-analysis. Clin. Microbiol. Infect., 25: 437-444.
Office International des Épizooties (OIE-World Organisation for Animal Health) (2018). Manual of diagnostic tests and vaccines for terrestrial animals, OIE. Paris. Chapter 3.1.12. PP: 503-516.
Priya, CG; Bhavani, K; Rathinam, SR and Muthukkaruppan, VR (2003). Identification and evaluation of LPS antigen for serodiagnosis of uveitis associated with Leptospirosis. J. Med. Microbiol., 52: 667-673.
Radostits, OM; Gay, CC; Hinchcliff, KW and Constable, PD (2010). Veterinary Medicine: A text book of the diseases of Cattle, Sheep, Pigs, Goats and Horses. 10th Edn., Edinburgh, London, New York, Oxford, Philadelphia, St. Louis, Sydney, Toronto, W. B. Saunders Elsevier. pp: 1102-1103.
Rani Prameela, D; Sreenivasulu, D; Vijayachari, P and Natarajseenivasan, K (2013). Seroepidemiology of leptospirosis in AndhraPradesh. Arch. Clin. Microbiol., 4: 1-10.
Sehgal, SC; Vijayachari, P; Sugunan, AP and Umapathi, T (2003). Field application of Lepto lateral flow for rapid diagnosis of Leptospirosis. J. Med. Microbiol., 52: 897-901.
Senthilkumar, K; Ravikumar, G and Aravindbabu, RP (2021). Spatio-temporal distribution of bovine leptospirosis in Tamil Nadu and a risk factor analysis. Vet. Med. (Praha), 66: 503-512.
Smits, HL; Eapen, CK; Sugathan, S; Kuriakose, M; Gasem, MH; Yersin, C; Sasaki, D; Pujianto, B; Vestering, M; Abdoel, TH and Gussenhoven, GC (2001). Lateral-Flow assay for rapid serodiagnosis of human Leptospirosis. Clin. Diagn. Lab. Immunol., 8: 166-169.
Srivastava, SK; Singh, SP and Srivastava, NC (1983). Sero-prevalence of leptospirosis in animals and man in India. Ind. J. Comp. Microbiol. Immunol. Infect. Dis., 4: 243-245.
Tansuphasiri, U; Deepradit, S; Phulsuksombati, D and Tangkanakul, W (2005). A test strip IgM Dot-ELISA
assay using Leptospiral antigen of endemic strains for serodiagnosis of acute Leptospirosis. J. Med. Assoc. Thail., 88: 391-398.
Terpstra, WJ; Ligthart, GS and Schoone, GJ (1985). ELISA for the detection of specific IgM and IgG in human leptospirosis. J. Gen. Microbiol., 131: 377-385.
Vanithamani, S; Shanmughapriya, S; Narayanan, R; Raja, V; Kanagavel, M; Sivasankari, K and Natarajaseenivasan, K (2013). Lipopolysaccharide specific immunochromatography based lateral flow assay for serogroup specific diagnosis of leptospirosis in India. Plos One. 10: e0137130.
Vijayachari, P (2007). Leptospira, In: Leptospirosis laboratory manual, Regional Medical Research Centre, Indian Council of Medical Research, Port Blair, India. https://apps.who.int/ iris/bitstream/handle/10665/205429/B2147.pdf?sequence=1.