Abstract
Listeria monocytogenes is a food-borne bacterium that causes listeriosis upon the ingestion of contaminated food. Traditional methods to detect L. monocytogenes require pre-enrichment broths to increase its concentration. To improve the screening of contaminated food and prevent listeriosis outbreaks, rapid, specific and sensitive assays are needed to detect L. monocytogenes. This study developed a prototype lateral flow immunochromatographic assay (LFIA) employing antibodies against L. monocytogenes Internalin A (InlA) and Internalin B (InlB) proteins, that are involved in non-phagocytic cell invasion. The following antibodies were used to capture L. monocytogenes antigenic targets: mouse anti-Internalin A monoclonal antibody (MAb-2D12) conjugated to colloidal gold nanoparticles and a mouse anti-Internalin B polyclonal antibody. This test was able to detect pure L. monocytogenes from culture with a limit of detection (LOD) ranging from 5.9 × 103 to 1.5 × 104 CFU/mL. In milk artificially contaminated with L. monocytogenes, the LOD was 1 × 105 CFU/mL. This prototype test discriminated L. monocytogenes from other bacterial species (Listeria innocua, Enterobacter cloacae, Bacillus cereus). Results indicate that this LFIA developed using antibodies against L. monocytogenes InlA and InlB proteins is a sensitive and specific tool that can be potentially useful to rapidly detect L. monocytogenes in contaminated food.
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Abbreviations
- AuNP:
-
Colloidal Gold Nanoparticle
- BHI:
-
Brain Heart Infusion
- InlA:
-
Internalin A
- InlB:
-
Internalin B
- LFIA:
-
Lateral Flow Immunochromatographic Assay
- LM:
-
Listeria Monocytogenes
- LOD:
-
Limit of Detection
- MAb:
-
Monoclonal Antibody
- NC:
-
Nitrocellulose Membrane
- OD:
-
Optical Density
- PAb:
-
Polyclonal Antibody
- TSB:
-
Tryptic Soy Broth
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Acknowledgements
Students’ fellowships were granted by: the Brazilian National Research Council (grant # 133188/2016-7 2016-2018), the Brazilian Coordination for the Improvement of Higher Education Personnel (CAPES grant # 88882.385457/2007-01 2019-2023) and the Alliances Program for Education and Training (Scholarships Brazil—PAEC OAS-GCUB). AK is a research fellow from the Brazilian Research Council/CNPq (“Bolsista de Produtividade em Pesquisa, Grant # 314366/2020-2). MMAS is PVN-II research fellow from FAPEAM/Amazonas (PECTI-AM/SAÚDE Program, Grant #004/2020) and a research fellow from the Brazilian Research Council/CNPq (“Bolsista de Produtividade em Pesquisa, Grant # 311986-2019-6).
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Conception and design: RSM, ESF, MM, FRC, MMAS, SBS; Acquisition of data: RSM, ESF, AV, DRS, ANM, TGS; Analysis and interpretation of data: LLL, ESF, RSM, AK, MMAS, SBS; Drafting the manuscript: LLL, ESF, SBS, MMAS; Revising the manuscript critically: LLL, RSM, AK, MM, FRC, MMAS, SBS; Approval of the final manuscript: LLL, ESF, MM, ANM, AV, DRS, TGS, RSM, MMAS, FRC, AK, SBS.
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Lopes-Luz, L., Silva-Filho, E., Mendonça, M. et al. Combined antibodies against internalins A and B proteins have potential application in immunoassay for detection of Listeria monocytogenes. J Food Sci Technol 60, 123–131 (2023). https://doi.org/10.1007/s13197-022-05597-9
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DOI: https://doi.org/10.1007/s13197-022-05597-9