Development and evaluation of polyclonal antibody-based antigen detection ELISA and dot blot assays as a less costly diagnostic alternative for pathogenic Leptospira infection

Tomckowiack, Camilo; Ramirez-Reveco, Alfredo; Henrriquez, Claudio; Salgado, Miguel

doi:10.1016/j.actatropica.2022.106782

Cited by 3 publications

(1 citation statement)

References 19 publications

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“…Leptospirosis continues to jeopardize the health of humans, livestock, and wild animals, causing economic losses and death. Diagnosis of the disease is usually based on serological testing with MAT; however, this test uses live cultures of various Leptospira serovars and specialized personnel, posing a challenge to field application [ 31 ]. Serological tests using recombinant antigens have higher sensitivity and specificity compared to whole bacterial cells because of the purity of the antigen and the reduction of non-specific parts in whole bacterial cells [ 32 ].…”

Section: Discussionmentioning

confidence: 99%

Production and characterization of immunoglobulin G anti-rLipL32 antibody as a biomarker for the diagnosis of leptospirosis

Susanti,

Sudarmono,

Dharmayanti

et al. 2024

Vet World

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Background and Aim: Microscopic agglutination test (MAT) for the diagnosis of leptospirosis requires live cultures and is serovar-specific, while polymerase chain reaction (PCR) requires expensive equipment and sample preparation. The rLipL32 protein is conserved and can be used for the production of immunoglobulin G (IgG) anti-rLipL32 antibody, which can be used as a biomarker for leptospirosis diagnosis. This study aimed to produce and characterize an IgG anti-rLipL32 antibody as a biomarker for leptospirosis diagnosis. Materials and Methods: Escherichia coli rLipL32 was cultured and analyzed by PCR and sequencing. Cultures were used for rLipL32 protein expression and purification and the rLipL32 protein was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The rLipL32 protein was used to produce anti-rLipL32 serum and was analyzed by enzyme-linked immunosorbent assay (ELISA). Serum was purified to obtain IgG anti-rLipL32 antibody and characterized by SDS-PAGE and western blotting. Results: PCR was able to amplify the LipL32 gene from E. coli rLipL32, and sequencing analysis showed 99.19% similarity with pathogenic Leptospira. SDS-PAGE analysis showed a 32-kDa band. ELISA results showed an increase in OD in anti-rLipL32 serum compared to preimmune serum. Western blotting results showed that the IgG anti-rLipL32 antibody was able to bind and cross-reacts with pathogenic Leptospira serovar but not with E. coli or Staphylococcus aureus. Conclusion: IgG anti-rLipL32 antibody has high specificity and sensitivity against Leptospira pathogens. These findings suggest that IgG anti-rLipL32 antibody is a promising biomarker for the diagnosis of leptospirosis. Keywords: anti-rLipL32 serum, immunoglobulin G anti-rLipL32 antibody, Leptospira, rLipL32 protein.

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Section: Discussionmentioning

confidence: 99%

Production and characterization of immunoglobulin G anti-rLipL32 antibody as a biomarker for the diagnosis of leptospirosis

Susanti,

Sudarmono,

Dharmayanti

et al. 2024

Vet World

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Pathogenic Leptospira Detection in Environmental Contaminant Water Sources by Highly Performance Antibody Absorption Polystyrene Agglutinating Particles

Suwannin,

Polpanich,

Lebaz

et al. 2024

Part & Part Syst Charact

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Leptospirosis is a re‐emerging bacterial zoonotic disease that affects both humans and animals, with a significantly higher incidence in tropical and sub‐tropical regions. Disease control, epidemiology, and surveillance rely on a One Health approach, as accurate detection can be applied to humans, animals, and the environment. This study represents the first attempt to develop a method for detecting the pathogenic Leptospira santarosai serovar Shermani based on the latex agglutination reaction. The serological activity of the antibody is examined to achieve a high titer of antibody before adsorption onto polystyrene particles. Using a pH medium of 6.8–7.8, total antibody adsorption of up to 3 mg m−2 is achieved. Particle agglutination is observed after incubating the antibody‐adsorbed PS with leptospiral culture for 4 min, revealing a detection limit of 1.7 × 102 leptospires mL−1. Interestingly, the detection limit increased by 1000 times when observing agglutination using spectrophotometer. The test exhibits high specificity with Shermani and shows negligible cross‐agglutination with non‐pathogenic Leptospira and water‐borne bacteria. Agglutination testing in collected water samples from natural sources demonstrates a good correlation with culture technique. This simple and rapid leptospires agglutination detection method can be applied as a screening test in environmental, human, and animal specimens.

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Leptospira-specific immunoglobulin Y (IgY) is protective in infected hamsters

Lv,

Xie,

Diao

et al. 2024

Vaccine

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Development and evaluation of polyclonal antibody-based antigen detection ELISA and dot blot assays as a less costly diagnostic alternative for pathogenic Leptospira infection

Cited by 3 publications

References 19 publications

Production and characterization of immunoglobulin G anti-rLipL32 antibody as a biomarker for the diagnosis of leptospirosis

Production and characterization of immunoglobulin G anti-rLipL32 antibody as a biomarker for the diagnosis of leptospirosis

Pathogenic Leptospira Detection in Environmental Contaminant Water Sources by Highly Performance Antibody Absorption Polystyrene Agglutinating Particles

Leptospira-specific immunoglobulin Y (IgY) is protective in infected hamsters

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