|作者：||Wu Yue, Wu Xudan, Hou Jinxiu, Chen Xiongnan, Huang Xiaobo, Zhou Bin|
1MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, China.
2College of Veterinary Medicine, Yangzhou University, Yangzhou, 225009, China.
3College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, 611130, China.
4MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, China. email@example.com.
|刊名：||BMC veterinary research, 2020, Vol.16 (1), pp.57|
|关键词：||Antibody detection; Clinical serum; Porcine parvovirus (PPV); Protein chip;|
|原始语种摘要：||BACKGROUND(#br)PPV is one of the most important pathogens causing porcine reproductive disorder. It has been shown in clinical cases to be a commonly mixed infection with other important swine diseases which can aggravate the severity of the disease and bring serious economic losses to the pig industry. Serological methods, such as hemagglutination inhibition assays (HAI), serum neutralization (SN), and the modified direct complement-fixation (MDCF) test were utilized earlier, whereas the enzyme-linked immunosorbent assay (ELISA) is the most frequently applied assay to detect PPV-specific antibodies.(#br)RESULTS(#br)We establish the visible protein chip and the cyanine dye 3 (Cy3)-labeled protein chip to detect the clinical serum from pigs. In this study, the recombinant protein VP2 of... PPV was expressed in E.coli, purified with nickel magnetic beads, and then printed onto epoxy-coated glass slides for preparation of the protein chip. After a series of experiments, the conditions of antigen protein concentration, incubation time of primary antibody or secondary antibody, and optimal serum dilution fold were optimized, resulting in a successful visible protein chip and Cy3-labeled protein chip. The results showed that the positive serum, diluted up to 6000-fold, can be detected by the visible protein chip, and the positive serum, diluted up to 12,800-fold, can be detected by the Cy3-labeled protein chip, suggesting the high sensitivity of these protein chips. Moreover, the positive detection ratio, sensitivity, and specificity of these two kinds of protein chips were higher than those of commercial ELISA antibody detection kits.(#br)CONCLUSION(#br)Overall, these two protein chips can be used to rapidly diagnose clinical samples with high throughput.|