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

Research (Published online: 08-04-2023)

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

Héctor D. Nájera-Rivera, Ana D. Rodríguez-Cortez, María G. Anaya-Santillán, Efrén Díaz-Aparicio, Ariadna V. Ramos-Rodríguez, Irlanda J. Siliceo-Cantero, Norma C. Vázquez-Franco, Erik Nieto-Patlán, Alejandro De Las Peñas, Liliana M. Valdés-Vázquez, and Laura Cobos-Marín

Veterinary World, 16(4): 704-710

Héctor D. Nájera-Rivera: Department of Reproduction, Veterinary School, UNAM, Mexico City, Mexico.

Ana D. Rodríguez-Cortez: Department of Reproduction, Veterinary School, UNAM, Mexico City, Mexico.

María G. Anaya-Santillán: Department of Microbiology and Immunology, Veterinary School, UNAM, Mexico City, Mexico.

Efrén Díaz-Aparicio: CENID, INIFAP, Mexico City, Mexico.

Ariadna V. Ramos-Rodríguez: Department of Microbiology and Immunology, Veterinary School, UNAM, Mexico City, Mexico.

Irlanda J. Siliceo-Cantero: Department of Microbiology and Immunology, Veterinary School, UNAM, Mexico City, Mexico.

Norma C. Vázquez-Franco: Division of Molecular Biology, IPICYT, San Luis Potosi, Mexico.

Erik Nieto-Patlán: Department of Microbiology and Immunology, Veterinary School, UNAM, Mexico City, Mexico.

Alejandro De Las Peñas: Division of Molecular Biology, IPICYT, San Luis Potosi, Mexico.

Liliana M. Valdés-Vázquez: Department of Microbiology and Immunology, Veterinary School, UNAM, Mexico City, Mexico.

Laura Cobos-Marín: Department of Microbiology and Immunology, Veterinary School, UNAM, Mexico City, Mexico.

doi: 10.14202/vetworld.2023.704-710

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

Corresponding author: Laura Cobos-Marín

E-mail: lcobosm@fmvz.unam.mx

Citation: 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, Mycobacterium avium subsp. Paratuberculosis, and Brucella melitensis in goats, Veterinary World, 16(4): 704-710.

Abstract

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.