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Expression of recombinant Leptospiral surface Lipoprotein-Lsa27 in E. coli and its evaluation for serodiagnosis of bovine leptospirosis by latex agglutination test.

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Title Expression of recombinant Leptospiral surface Lipoprotein-Lsa27 in E. coli and its evaluation for serodiagnosis of bovine leptospirosis by latex agglutination test.
Not Available
 
Creator Alamuri A
Kumar KV
Sowjanya KS
Linshamol L
Sridevi R
Nagalingam M
Roy P
Balamurugan V
 
Subject Leptospira
Lsa27 (LIC12895)
Recombinant protein
Latex agglutination test
Bovine
Serodiagnosis
 
Description Not Available
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.
Not Available
 
Date 2021-04-19T09:13:53Z
2021-04-19T09:13:53Z
2020-10-02
 
Type Research Paper
 
Identifier Alamuri A, Kumar KV, Sowjanya KS, Linshamol L, Sridevi R, Nagalingam M, Roy P and Balamurugan V. (2020). Expression of recombinant Leptospiral surface Lipoprotein-Lsa27 in E. coli and its evaluation for serodiagnosis of bovine leptospirosis by latex agglutination test. Molecular Biotechnology. 62(11): 598-610.
1073-6085 (print)
1559-0305 (web)
http://krishi.icar.gov.in/jspui/handle/123456789/46519
 
Language English
 
Relation Not Available;
 
Publisher Springer Science+Business Media