Giménez-Lirola Luis G, Jiang Yong-Hou, Sun Dong, Hoang Hai, Yoon Kyoung-Jin, Halbur Patrick G, Opriessnig Tanja
Department of Veterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, Iowa, USA.
Clin Vaccine Immunol. 2014 Jan;21(1):85-95. doi: 10.1128/CVI.00451-13. Epub 2013 Nov 13.
Surveillance for the presence of Actinobacillus pleuropneumoniae infection in a population plays a central role in controlling the disease. In this study, a 4-plex fluorescent microbead-based immunoassay (FMIA), developed for the simultaneous detection of IgG antibodies to repeat-in-toxin (RTX) toxins (ApxI, ApxII, ApxIII, and ApxIV) of A. pleuropneumoniae, was evaluated using (i) blood serum samples from pigs experimentally infected with each of the 15 known A. pleuropneumoniae serovars or with Actinobacillus suis, (ii) blood serum samples from pigs vaccinated with a bacterin containing A. pleuropneumoniae serovar 1, 3, 5, or 7, and (iii) blood serum samples from pigs with an unknown A. pleuropneumoniae exposure status. The results were compared to those obtained in a previous study where a dual-plate complement fixation test (CFT) and three commercially available enzyme-linked immunosorbent assays (ELISAs) were conducted on the same sample set. On samples from experimentally infected pigs, the 4-plex Apx FMIA detected specific seroconversion to Apx toxins as early as 7 days postinfection in a total of 29 pigs inoculated with 14 of the 15 A. pleuropneumoniae serovars. Seroconversion to ApxII and ApxIII was detected by FMIA in pigs inoculated with A. suis. The vaccinated pigs showed poor humoral responses against ApxI, ApxII, ApxIII, and ApxIV. In the field samples, the humoral response to ApxIV and the A. pleuropneumoniae seroprevalence increased with age. This novel FMIA (with a sensitivity of 82.7% and a specificity of 100% for the anti-ApxIV antibody) was found to be more sensitive and accurate than current tests (sensitivities, 9.5 to 56%; specificity, 100%) and is potentially an improved tool for the surveillance of disease and for monitoring vaccination compliance.
监测群体中胸膜肺炎放线杆菌感染的情况在控制该疾病方面起着核心作用。在本研究中，一种基于四重荧光微珠的免疫测定法（FMIA）被开发用于同时检测针对胸膜肺炎放线杆菌重复毒素（RTX）毒素（ApxI、ApxII、ApxIII和ApxIV）的IgG抗体，并使用以下样本进行了评估：（i）来自经15种已知胸膜肺炎放线杆菌血清型中的每一种或猪放线杆菌实验感染的猪的血清样本；（ii）来自接种了含有胸膜肺炎放线杆菌血清型1、3、5或7的菌苗的猪的血清样本；（iii）来自胸膜肺炎放线杆菌暴露状态未知的猪的血清样本。将结果与先前一项研究的结果进行了比较，在先前的研究中，对同一组样本进行了双板补体结合试验（CFT）和三种市售酶联免疫吸附测定（ELISA）。在来自实验感染猪的样本中，四重Apx FMIA在总共29头接种了15种胸膜肺炎放线杆菌血清型中的14种的猪中，最早在感染后7天就检测到了针对Apx毒素的特异性血清转化。在接种猪放线杆菌的猪中，FMIA检测到了针对ApxII和ApxIII的血清转化。接种疫苗的猪对ApxI、ApxII、ApxIII和ApxIV的体液反应较差。在现场样本中，对ApxIV的体液反应和胸膜肺炎放线杆菌血清阳性率随年龄增加。这种新型FMIA（抗ApxIV抗体的灵敏度为82.7%，特异性为100%）被发现比目前的检测方法（灵敏度为9.5%至56%；特异性为100%）更灵敏、准确，并且可能是一种用于疾病监测和监测疫苗接种依从性的改进工具。
