Indirect ELISA (iELISA) for routine detection of antibodies against Minute Virus of Mice (MVM) in mice colonies

• Published in: Revista argentina de microbiología 2017-07, Vol.49 (3), p.210-215
• Main Authors: Laborde, Juan M., Sguazza, Guillermo H., Fuentealba, Nadia A., Corva, Santiago G., Carbone, Cecilia, Galosi, Cecilia M.
• Format: Article
• Language: English
• Subjects: Animals; Antibodies, Viral - analysis; Diagnosis; Diagnóstico; ELISA; Enzyme-Linked Immunosorbent Assay; Fluorescent Antibody Technique, Indirect; Mice; Minute Virus of Mice; Minute Virus of Mice - immunology; Reproducibility of Results; Sensitivity and Specificity; Virus diminuto del ratón
• ISSN: 0325-7541
• DOI: 10.1016/j.ram.2017.02.005

In this study we developed an indirect ELISA to detect antibodies against Minute Virus of Mice (MVM) using an antigen produced from BHK-21 cells infected with a prototype strain of the virus. The optimal antigen concentration and serum dilutions were established. In order to analyze variability in the laboratory, reproducibility and repeatability within and between plates were determined. Then, a panel of 460 sera from conventional facilities and previously classified as positive or negative by the indirect fluorescent antibody assay was analyzed. The cutoff value was determined by a receiver operating characteristic (ROC) curve. The results of the indirect ELISA were compared with those of the indirect fluorescent antibody assay. The ELISA assay showed 100% sensitivity and 99% specificity. ELISA is a useful tool to be developed in standard virology laboratories and can be used for screening animals faster than the traditional indirect fluorescent antibody assay. Se desarrolló un ELISA indirecto para detectar anticuerpos contra el virus diminuto del ratón (Mice minute virus [MVM]), utilizando un antígeno producido a partir de células BHK-21 infectadas con la cepa prototipo del virus. Se establecieron las diluciones óptimas de antígeno y el suero a utilizar. Para analizar la variabilidad en el laboratorio, se determinaron la reproducibilidad y la repetibilidad dentro de una placa y entre placas. Luego se analizaron 460 sueros provenientes de bioterios convencionales y clasificados previamente como positivos o negativos por inmunofluorescencia indirecta. El valor de corte se determinó mediante una curva ROC. Los resultados se compararon con los obtenidos con la prueba de inmunofluorescencia indirecta. El ELISA mostró 100% de sensibilidad y un 99% de especificidad. Esta técnica demostró ser una herramienta útil para desarrollar en laboratorios de virología estándar y puede utilizarse como prueba tamiz para seleccionar animales de manera más rápida que con la tradicional prueba de inmunofluorescencia indirecta.