Institute for Food Quality and Food Safety, University of Veterinary Medicine Hannover, Foundation, Bischofsholer Damm 15, D-30173, Hannover, Germany.
Abbott (Alere Technologies GmbH), Löbstedter Straße 103-105, D-07749, Jena, Germany.
BMC Vet Res. 2020 Apr 6;16(1):106. doi: 10.1186/s12917-020-02308-4.
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.
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.
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.
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.
屠宰场对猪群进行血清学筛查被认为是改进肉品检验程序和畜群健康管理的潜在工具。因此,我们之前报道了一种微型蛋白质微阵列作为一种新的血清 IgG 筛查试验,用于检测猪的人畜共患病原体和生产疾病的可行性。本研究调查了基于蛋白质微阵列的检测方法是否适用于使用血清或肉汁进行高通量样本检测。
雅培(前身为 Alere 技术公司)德国耶拿生产的带有 12 种不同抗原的微阵列分别在之前提供的“ArrayTube”平台和用于大规模使用的“ArrayStrip”平台上制作。开发了一种用于两种微阵列平台的肉汁检测方案。使用来自德国三个屠宰场的 88 对配对样本分析血清和肉汁之间的一致性。血清稀释 1:50,肉汁稀释 1:2。使用前一项研究的所有测试抗原的 ELISA 结果作为参考测试,在两种微阵列平台上对两种测试标本进行受试者工作特征分析。
使用血清作为测试标本时,对弓形虫(0.87)、小肠结肠炎耶尔森菌(0.97)、猪肺炎支原体(0.84)和胸膜肺炎放线杆菌(0.71)的分析得出了较高的曲线下面积值(AUC>0.7),而使用肉汁作为测试标本时,对弓形虫(0.99)、小肠结肠炎耶尔森菌(0.94)、猪繁殖与呼吸综合征病毒(0.88)、胸膜肺炎放线杆菌(0.78)和沙门氏菌属(0.72)的分析得出了较高的 AUC 值。在 ArrayStrip 平台上,对血清和肉汁进行比较时,弓形虫的 Cohens kappa 值为 0.92,小肠结肠炎耶尔森菌的 Cohens kappa 值为 0.82。当在另外两个实验室应用新方法时,对于这两种病原体,实验室之间的 kappa 值在 0.63 到 0.94 之间。
进一步开发微型猪特异性 IgG 蛋白质微阵列检测方法表明,肉汁可以在微阵列平台上使用。在 ArrayTube 和 ArrayStrip 平台上,使用两种样本材料,两种测试抗原(弓形虫、小肠结肠炎耶尔森菌)的检测准确性均较高。