Development of a Multispecies Double-Antigen Sandwich ELISA Using N and RBD Proteins to Detect Antibodies against SARS-CoV-2
SARS-CoV-2 infects humans and a broad spectrum of animal species, such as pets, zoo animals, and nondomestic animals. Monitoring infection in animals is important in terms of the risk of interspecies transmission and the emergence of new viral variants. Economical, fast, efficient, and sensitive dia...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-11-01
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| Series: | Animals |
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| Online Access: | https://www.mdpi.com/2076-2615/13/22/3487 |
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| author | Maritza Cordero-Ortiz Mónica Reséndiz-Sandoval Freddy Dehesa-Canseco Mario Solís-Hernández Jahir Pérez-Sánchez Carlos Martínez-Borges Verónica Mata-Haro Jesús Hernández |
| author_facet | Maritza Cordero-Ortiz Mónica Reséndiz-Sandoval Freddy Dehesa-Canseco Mario Solís-Hernández Jahir Pérez-Sánchez Carlos Martínez-Borges Verónica Mata-Haro Jesús Hernández |
| author_sort | Maritza Cordero-Ortiz |
| collection | DOAJ |
| description | SARS-CoV-2 infects humans and a broad spectrum of animal species, such as pets, zoo animals, and nondomestic animals. Monitoring infection in animals is important in terms of the risk of interspecies transmission and the emergence of new viral variants. Economical, fast, efficient, and sensitive diagnostic tests are needed to analyze animal infection. Double-antigen sandwich ELISA has the advantage of being multispecies and can be used for detecting infections caused by pathogens that infect several animal hosts. This study aimed to develop a double-antigen sandwich ELISA using two SARS-CoV-2 proteins, N and RBD. We compared its performance, when using these proteins separately, with an indirect ELISA and with a surrogate virus neutralization test. Positive and negative controls from a cat population (<i>n</i> = 31) were evaluated to compare all of the tests. After confirming that double-antigen sandwich ELISA with both RBD and N proteins had the best performance (AUC= 88%), the cutoff was adjusted using positive and negative samples from cats, humans (<i>n</i> = 32) and guinea pigs (<i>n</i> = 3). The use of samples from tigers (<i>n</i> = 2) and rats (<i>n</i> = 51) showed good agreement with the results previously obtained using the microneutralization test. Additionally, a cohort of samples from dogs with unknown infection status was evaluated. These results show that using two SARS-CoV-2 proteins in the double-antigen sandwich ELISA increases its performance and turns it into a valuable assay with which to monitor previous infection caused by SARS-CoV-2 in different animal species. |
| first_indexed | 2024-03-09T17:05:42Z |
| format | Article |
| id | doaj.art-abc9eb3719104ee5b11325c9b9df973e |
| institution | Directory Open Access Journal |
| issn | 2076-2615 |
| language | English |
| last_indexed | 2024-03-09T17:05:42Z |
| publishDate | 2023-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Animals |
| spelling | doaj.art-abc9eb3719104ee5b11325c9b9df973e2023-11-24T14:24:42ZengMDPI AGAnimals2076-26152023-11-011322348710.3390/ani13223487Development of a Multispecies Double-Antigen Sandwich ELISA Using N and RBD Proteins to Detect Antibodies against SARS-CoV-2Maritza Cordero-Ortiz0Mónica Reséndiz-Sandoval1Freddy Dehesa-Canseco2Mario Solís-Hernández3Jahir Pérez-Sánchez4Carlos Martínez-Borges5Verónica Mata-Haro6Jesús Hernández7Laboratorio de Inmunología, Centro de Investigación en Alimentación y Desarrollo, A.C., Hermosillo 83304, Sonora, MexicoLaboratorio de Inmunología, Centro de Investigación en Alimentación y Desarrollo, A.C., Hermosillo 83304, Sonora, MexicoComisión México-Estados Unidos para la Prevención de la Fiebre Aftosa y otras Enfermedades Exóticas de los Animales (CPA), Servicio Nacional de Sanidad, Inocuidad y Calidad Agroalimentaria (SENASICA), Secretaría de Agricultura y Desarrollo Rural (SADER), Ciudad de Mexico 05110, Mexico State, MexicoComisión México-Estados Unidos para la Prevención de la Fiebre Aftosa y otras Enfermedades Exóticas de los Animales (CPA), Servicio Nacional de Sanidad, Inocuidad y Calidad Agroalimentaria (SENASICA), Secretaría de Agricultura y Desarrollo Rural (SADER), Ciudad de Mexico 05110, Mexico State, MexicoCentro de Biotecnología Genómica, Instituto Politécnico Nacional, Cd., Reynosa 88710, Tamaulipas, MexicoHospital Veterinario Borges, Hermosillo 83600, Sonora, MexicoLaboratorio de Microbiología e Inmunología, Centro de Investigación en Alimentación y Desarrollo, A.C., Hermosillo 83304, Sonora, MexicoLaboratorio de Inmunología, Centro de Investigación en Alimentación y Desarrollo, A.C., Hermosillo 83304, Sonora, MexicoSARS-CoV-2 infects humans and a broad spectrum of animal species, such as pets, zoo animals, and nondomestic animals. Monitoring infection in animals is important in terms of the risk of interspecies transmission and the emergence of new viral variants. Economical, fast, efficient, and sensitive diagnostic tests are needed to analyze animal infection. Double-antigen sandwich ELISA has the advantage of being multispecies and can be used for detecting infections caused by pathogens that infect several animal hosts. This study aimed to develop a double-antigen sandwich ELISA using two SARS-CoV-2 proteins, N and RBD. We compared its performance, when using these proteins separately, with an indirect ELISA and with a surrogate virus neutralization test. Positive and negative controls from a cat population (<i>n</i> = 31) were evaluated to compare all of the tests. After confirming that double-antigen sandwich ELISA with both RBD and N proteins had the best performance (AUC= 88%), the cutoff was adjusted using positive and negative samples from cats, humans (<i>n</i> = 32) and guinea pigs (<i>n</i> = 3). The use of samples from tigers (<i>n</i> = 2) and rats (<i>n</i> = 51) showed good agreement with the results previously obtained using the microneutralization test. Additionally, a cohort of samples from dogs with unknown infection status was evaluated. These results show that using two SARS-CoV-2 proteins in the double-antigen sandwich ELISA increases its performance and turns it into a valuable assay with which to monitor previous infection caused by SARS-CoV-2 in different animal species.https://www.mdpi.com/2076-2615/13/22/3487SARS-CoV-2COVID-19ELISAmicroneutralizationcatsmultispecies |
| spellingShingle | Maritza Cordero-Ortiz Mónica Reséndiz-Sandoval Freddy Dehesa-Canseco Mario Solís-Hernández Jahir Pérez-Sánchez Carlos Martínez-Borges Verónica Mata-Haro Jesús Hernández Development of a Multispecies Double-Antigen Sandwich ELISA Using N and RBD Proteins to Detect Antibodies against SARS-CoV-2 Animals SARS-CoV-2 COVID-19 ELISA microneutralization cats multispecies |
| title | Development of a Multispecies Double-Antigen Sandwich ELISA Using N and RBD Proteins to Detect Antibodies against SARS-CoV-2 |
| title_full | Development of a Multispecies Double-Antigen Sandwich ELISA Using N and RBD Proteins to Detect Antibodies against SARS-CoV-2 |
| title_fullStr | Development of a Multispecies Double-Antigen Sandwich ELISA Using N and RBD Proteins to Detect Antibodies against SARS-CoV-2 |
| title_full_unstemmed | Development of a Multispecies Double-Antigen Sandwich ELISA Using N and RBD Proteins to Detect Antibodies against SARS-CoV-2 |
| title_short | Development of a Multispecies Double-Antigen Sandwich ELISA Using N and RBD Proteins to Detect Antibodies against SARS-CoV-2 |
| title_sort | development of a multispecies double antigen sandwich elisa using n and rbd proteins to detect antibodies against sars cov 2 |
| topic | SARS-CoV-2 COVID-19 ELISA microneutralization cats multispecies |
| url | https://www.mdpi.com/2076-2615/13/22/3487 |
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