Multiplex assay for the simultaneous detection of antibodies against small ruminant lentivirus, Mycobacterium avium subsp. paratuberculosis, and Brucella melitensis in goats

Vet World Vol.16 April-2023 Article-6

Research Article

Veterinary World, 16(4): 704-710

https://doi.org/10.14202/vetworld.2023.704-710

Héctor D. Nájera-Rivera¹, Ana D. Rodríguez-Cortez¹, María G. Anaya-Santillán², Efrén Díaz-Aparicio³, Ariadna V. Ramos-Rodríguez², Irlanda J. Siliceo-Cantero², Norma C. Vázquez-Franco⁴, Erik Nieto-Patlán², Alejandro De Las Peñas⁴, Liliana M. Valdés-Vázquez², and Laura Cobos-Marín²

1. Department of Reproduction, Veterinary School, UNAM, Mexico City, Mexico.
2. Department of Microbiology and Immunology, Veterinary School, UNAM, Mexico City, Mexico.
3. CENID, INIFAP, Mexico City, Mexico.
4. Division of Molecular Biology, IPICYT, San Luis Potosi, Mexico.

Background and Aim: Brucellosis, paratuberculosis (PTb), and infections caused by small ruminant lentivirus (SRLV), formerly known as caprine arthritis encephalitis virus (CAEV), adversely affect goat production systems. Nonetheless, commonly used diagnostic tests can only determine one analyte at a time, increasing disease surveillance costs, and limiting their routine use. This study aimed to design and validate a multiplex assay for antibody detection against these three diseases simultaneously.

Materials and Methods: Two recombinant proteins from the SRLV (p16 and gp38), the native hapten of Brucella melitensis, and the paratuberculosis-protoplasmic antigen 3 from Mycobacterium avium subsp. paratuberculosis (MAP) were used to devise and assess a multiplex assay. Conditions for the Luminex® multiplex test were established and validated by sensitivity, specificity, repeatability, and reproducibility parameters. Cut-off points for each antigen were also established.

Results: The 3-plex assay had high sensitivity (84%) and specificity (95%). The maximum coefficients of variation were 23.8% and 20.5% for negative and positive control samples, respectively. The p16 and gp38 SRLV antigens are 97% and 95%, similar to the CAEV sequence found in GenBank, respectively.

Conclusion: The multiplex test can be effectively used for the simultaneous detection of antibodies against SRLV, MAP and B. melitensis in goats. Keywords: antibody detection, brucellosis, Luminex®, paratuberculosis, serological test, small ruminant lentivirus.

Keywords: antibody detection, brucellosis, Luminex®, paratuberculosis, serological test, small ruminant lentivirus.

How to cite this article: Nájera-Rivera HD, Rodríguez-Cortez AD, Anaya-Santillán MG, Díaz-Aparicio E, Ramos-Rodríguez AV, Siliceo-Cantero IJ, Vázquez-Franco NC, Nieto-Patlán E, De Las Peñas A, Valdés-Vázquez LM, and Cobos-Marín L (2023) Multiplex assay for the simultaneous detection of antibodies against small ruminant lentivirus,

Received: 07-11-2022 Accepted: 27-02-2023 Published online: 08-04-2023

Corresponding author: Laura Cobos-Marín E-mail: lcobosm@fmvz.unam.mx

DOI: 10.14202/vetworld.2023.704-710

Copyright: Nájera-Rivera, et al. This article is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http:// creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.