Choi K H, Maheswaran S K, Molitor T W
Department of Veterinary Pathobiology, University of Minnesota, St. Paul 55108.
Avian Dis. 1990 Jul-Sep;34(3):539-47.
A dot immunobinding assay (DIA) was developed to detect antibodies against Pasteurella multocida in turkey serum. Five coating antigens, namely, whole-cell (WC) antigen, sonicated cell lysate (SCL), crude capsular extract (CE), formalin extract (FE), and heat-stable antigen (HSA), were compared by enzyme-linked immunosorbent assay (ELISA) and DIA using reference antisera against P. multocida organisms. WC and SCL antigens showed higher sensitivity, whereas FE and HSA antigens were more specific coating antigens in both assays. The specificity of DIA was greater than ELISA by comparing the P/N ratios of HSA against serum prepared from heterologous serotype of P. multocida. The DIA had also several distinct advantages over the ELISA, which included reduction of the manipulation time and more uniform binding of coating antigens onto the nitrocellulose membranes compared with binding of coating antigens to microtiter plates for ELISA.
开发了一种斑点免疫结合测定法(DIA)以检测火鸡血清中抗多杀巴斯德菌的抗体。使用针对多杀巴斯德菌的参考抗血清,通过酶联免疫吸附测定法(ELISA)和DIA比较了五种包被抗原,即全细胞(WC)抗原、超声破碎细胞裂解物(SCL)、粗荚膜提取物(CE)、福尔马林提取物(FE)和热稳定抗原(HSA)。WC和SCL抗原显示出更高的灵敏度,而FE和HSA抗原在两种测定中都是更具特异性的包被抗原。通过比较HSA与由多杀巴斯德菌异源血清型制备的血清的P/N比值,DIA的特异性高于ELISA。与ELISA相比,DIA还具有几个明显的优势,包括减少操作时间以及与包被抗原与ELISA微量滴定板的结合相比,包被抗原在硝酸纤维素膜上的结合更均匀。