Use of meat juice and blood serum with a miniaturised protein microarray assay to develop a multi-parameter IgG screening test with high sample throughput potential for slaughtering pigs

Abstract

Background: Serological screening of pig herds at the abattoir is considered a potential tool to improve meat inspection procedures and herd health management. Therefore, we previously reported the feasibility of a miniaturised protein microarray as a new serological IgG screening test for zoonotic agents and production diseases in pigs. The present study investigates whether the protein microarray-based assay is applicable for high sample throughput using either blood serum or meat juice.Material and methods: Microarrays with 12 different antigens were produced by Abbott (formerly Alere Technologies GmbH) Jena, Germany in a previously offered 'ArrayTube' platform and in an 'ArrayStrip' platform for large-scale use. A test protocol for the use of meat juice on both microarray platforms was developed. Agreement between serum and meat juice was analysed with 88 paired samples from three German abattoirs. Serum was diluted 1:50 and meat juice 1:2. ELISA results for all tested antigens from a preceding study were used as reference test to perform Receiver Operating Characteristic analysis for both test specimens on both microarray platforms.Results: High area under curve values (AUC > 0.7) were calculated for the analysis of T. gondii (0.87), Y. enterocolitica (0.97), Mycoplasma hyopneumoniae (0.84) and Actinobacillus pleuropneumoniae (0.71) with serum as the test specimen and for T. gondii (0.99), Y. enterocolitica (0.94), PRRSV (0.88), A. pleuropneumoniae (0.78) and Salmonella spp. (0.72) with meat juice as the test specimen on the ArrayStrip platform. Cohens kappa values of 0.92 for T. gondii and 0.82 for Y. enterocolitica were obtained for the comparison between serum and meat juice. When applying the new method in two further laboratories, kappa values between 0.63 and 0.94 were achieved between the laboratories for these two pathogens.(Continued on next page) Conclusion: Further development of a miniaturised pig-specific IgG protein microarray assay showed that meat juice can be used on microarray platforms. Two out of twelve tested antigens (T. gondii, Y. enterocolitica) showed high test accuracy on the ArrayTube and the ArrayStrip platform with both sample materials.

Development of an Immunochromatographic Test Based on Rhoptry Protein 14 for Serological Detection of Toxoplasma gondii Infection in Swine

“…A novel luciferase-linked antibody capture assay (LACA) [ 34 ] and a fluorescent bead-based multiplex assay [ 35 ] were recently established for the individually serological diagnosis of T. gondii in pigs and chickens. In addition, a miniaturized protein microarray assay was recently developed for analysis of antibodies in meat juice and blood serum with high sample throughput for the slaughter of pigs [ 36 ]. At present, most serologic methods require technical training and are laborious difficult to use at the point of care.…”

Estudios, detección y problemática de Toxoplasma Gondii en cerdos

“…gondii en animales. Los microarrays de proteínas podrían ser útiles en procesar para el análisis de anticuerpos en jugo de carne y suero para un alto rendimiento de muestras para animales sacrificados (Loreck et al, 2020).…”

Development of a droplet digital polymerase chain reaction tool for the detection of Toxoplasma gondii in meat samples

“…Moreover, new serological tools have been developed for the detection of T. gondii in chickens, using Luminex technology (Fabian et al, 2020), a luciferase-linked antibody capture assay (LACA) (Duong et al, 2020) or microarrays to detect antibodies in meat juice and serum (Loreck et al, 2020).…”