J Vet Sci.  2012 Mar;13(1):99-101. 10.4142/jvs.2012.13.1.99.

Evaluation and comparison of native and recombinant LipL21 protein-based ELISAs for diagnosis of bovine leptospirosis

• Joseph S
• Thomas N
• Thangapandian E
• Singh VP
• Verma R
• Srivastava SK

• KMID: 1365009
• DOI: http://doi.org/10.4142/jvs.2012.13.1.99

Abstract

A 21-kDa leptospiral lipoprotein (LipL21) was evaluated for its diagnostic potential to detect bovine leptospirosis by ELISA. Both native LipL21 (nLipL21) and recombinant LipL21 (rLipL21) proteins were tested and compared regarding diagnostic efficiency, and no statistically significant difference was observed. The sensitivity of rLipL21 ELISA for 62 microscopic agglutination test (MAT) positive sera was 100% and the specificity with 378 MAT negative sera was 97.09%. Thus, rLipL21 protein-based ELISA could be used as an alternative to MAT for the diagnosis of bovine leptospirosis.

Keyword

ELISA; leptospirosis; microscopic agglutination test; nLipL21; rLipL21

MeSH Terms

Animals
Antibodies, Bacterial/blood
Antigens, Bacterial/biosynthesis/*chemistry/genetics
Bacterial Outer Membrane Proteins/biosynthesis/*chemistry/genetics
Cattle
Cattle Diseases/blood/*microbiology
Enzyme-Linked Immunosorbent Assay/veterinary
Leptospira interrogans/*isolation & purification
Leptospirosis/blood/microbiology/*veterinary
Lipoproteins/biosynthesis/*chemistry/genetics
Recombinant Proteins/biosynthesis/chemistry/genetics
Sensitivity and Specificity

Figure

• 

  Fig. 1 Western blot analysis. Lane 1-3: checking the identity and immunogenicity of rLipL21 using nLipL21 hyperimmune serum, Lane 1: induced Escherichia (E.) coli cell lysate, Lane 2: non-induced E. coli cell lysate, Lane 3: purified rLipL21, Lanes 4-6: analysis of the immunoreactivity levels of rLipL21 and nLipL21 to MAT positive field cattle sera, Lane 4: purified rLipL21 protein, Lane 5: sarcosyl-soluble portion of OMP, Lane 6: sarcosyl-insoluble portion of OMP, Lanes 7-12: checking the cross-reactivity of rLipL21 hyperimmune sera to various bacteria, Lane 7: purified rLipL21, Lane 8: Brucella abortus culture lysate, Lane 9: Campylobacter jejuni culture lysate, Lane 10: Mycoplasma bovis culture lysate, Lane 11: Pasteurella multocida culture lysate, Lane 12: Salmonella typhimurium culture lysate.

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