Combined antibodies against internalins A and B proteins have potential application in immunoassay for detection of Listeria monocytogenes

Leonardo Lopes-Luz; Ernandes Silva-Filho; Marcelo Mendonça; Ângela Nunes Moreira; Andressa Venceslau; Dienny Rodrigues de Sousa; Tatiana Galvez Sánchez; Rodrigo Scaliante de Moura; Fabricio Rochedo Conceição; André Kipnis; Mariane Martins de Araújo Stefani; Samira Bührer-Sékula

doi:10.1007/s13197-022-05597-9

Combined antibodies against internalins A and B proteins have potential application in immunoassay for detection of Listeria monocytogenes

J Food Sci Technol. 2023 Jan;60(1):123-131. doi: 10.1007/s13197-022-05597-9. Epub 2022 Dec 6.

Affiliations

¹ Instituto de Patologia Tropical e Saúde Pública, Universidade Federal de Goiás, Goiânia, GO 74605-050 Brasil.
² Universidade Federal Do Agreste Pernambuco, Curso de Medicina Veterinária, Garanhuns, Pernambuco 55292-270 Brasil.
³ Centro de Desenvolvimento Tecnológico, Núcleo de Biotecnologia, Laboratório de Imunologia Aplicada, Universidade Federal de Pelotas, Pelotas, RS 96010-610 Brasil.
⁴ Associação Educativa Evangélica, Anápolis, GO 75085-230 Brasil.

^# Contributed equally.

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.

Supplementary information: The online version contains supplementary material available at 10.1007/s13197-022-05597-9.

Keywords: Anti-Internalin A monoclonal antibody; Anti-Internalin B polyclonal antibodies; Lateral flow immunoassay; Listeria monocytogenes; Listeriosis; Rapid test.

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