Expression of Recombinant Leptospiral Surface Lipoprotein-Lsa27 in E. coli and Its Evaluation for Serodiagnosis of Bovin
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ORIGINAL PAPER
Expression of Recombinant Leptospiral Surface Lipoprotein‑Lsa27 in E. coli and Its Evaluation for Serodiagnosis of Bovine Leptospirosis by Latex Agglutination Test Anusha Alamuri1,2 · K. Vinod Kumar1 · S. SowjanyaKumari1 · L. Linshamol1 · R. Sridevi1 · M. Nagalingam1 · Parimal Roy1 · V. Balamurugan1 Accepted: 24 September 2020 © Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract The expressed recombinant leptospiral surface adhesion lipoprotein (Lsa27) of pathogenic Leptospira in E. coli was evaluated for the detection of Leptospira antibodies in cattle sera by latex agglutination test (LAT). The Lsa27 lacking signal peptide coding gene sequences from L. interrogans serovar Pomona was amplified (~ 660 bp) by PCR and the amplicon was cloned into pETiteN-HisKan vector. The expressed recombinant Lsa27 histidine-tagged fusion protein (rLsa27) was Ni–NTA affinity purified under denaturation followed by renaturation methods. The purified rLsa27 was characterized by SDS-PAGE and immunoblot, which confirmed the leptospiral protein with a MW of ~ 25 kDa. Further, the prepared sensitized latex beads coated with rLsa27 were evaluated as a diagnostic antigen for detection of pathogenic Leptospira antibodies by using known microscopic agglutination test (MAT) positive (n = 74) and negative (n = 62) sera for Leptospira antibodies in LAT, which revealed the relative diagnostic sensitivity of 91.89% and specificity of 87.10% against the gold standard serological test, MAT. Furthermore, on evaluation of developed rLsa27 LAT using serum samples from cattle associated with the history of abortions and reproductive disorder (n = 309), the relative sensitivity of 96.15%, and specificity of 89.11% were observed. Therefore, this rapid field test using the rLsa27 is first of its kind and it could be used as a screening test for the detection of Leptospira antibodies or it can be complemented by other diagnostics for the diagnosis /surveillance of bovine leptospirosis. Keywords Leptospira · Lsa27 (LIC12895) · Recombinant protein · Latex agglutination test · Bovine · Serodiagnosis
Introduction Leptospirosis is an emerging and re-emerging neglected zoonotic disease with worldwide distribution and it is caused by serovars of the pathogenic species spirochetes belonging to the genus Leptospira [1]. It is a fast-emerging zoonosis of major public health concern affecting the health of both the domestic animals and humans in tropical and sub-tropical countries [2, 3]. The disease affects a variety of hosts including livestock [4] resulting in heavy economic losses * V. Balamurugan [email protected]; [email protected] 1
Indian Council of Agricultural Research -National Institute of Veterinary Epidemiology and Disease Informatics (ICAR-NIVEDI), Yelahanka, Post Box No. 6450, Bengaluru, Karnataka 560 064, India
Jain University, Bengaluru, Karnataka, India
2
to the farming community on account of reproductive disorders [5]. Leptospirosis in livestock especially in bovines causes repro
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