Johansson M E, Grandien M, Arro L
J Immunol Methods. 1979;27(3):263-72. doi: 10.1016/0022-1759(79)90223-0.
The conditions for preparation of type-specific and subtype-specific influenza A virus reagents to be used in the immunofluorescence technique have been evaluated. Type A-specific antibodies were prepared by passing an antivirion hyperimmune serum through an immunoadsorbent column containing antigens from disrupted virions of a different influenza A virus subtype. The type-specific antibodies were recovered from the immunoadsorbent by desorption with 3 M NaI. For subtype determination, antisera against the various hemagglutinins were used. Such sera could be prepared by removal of irrelevant influenza A virus antibodies from sera directed against purified virions and isolated peplomers, respectively. This was performed by passing the antisera through immunoadsorbent columns containing antigens from disrupted virions of appropriate strains. However, attempts to obtain a hemagglutin-specific antiserum from a serum directed against allantoic fluid virus suspension failed with this procedure. Antisera obtained after immunization with purified hemagglutinin were also elaborated. These sera were shown to be superior for subtyping of influenza A virus infections by immunofluorescence, but could not a priori be regarded as subtype-specific. The usefulness of subtype-specific sera has been demonstrated on clinical specimens for rapid virus diagnosis.
已对用于免疫荧光技术的甲型流感病毒型特异性和亚型特异性试剂的制备条件进行了评估。通过使抗病毒超免疫血清通过含有来自不同甲型流感病毒亚型的破碎病毒粒子抗原的免疫吸附柱来制备甲型特异性抗体。通过用3M NaI解吸从免疫吸附剂中回收型特异性抗体。对于亚型测定，使用针对各种血凝素的抗血清。此类抗血清可分别通过从针对纯化病毒粒子和分离的纤突的血清中去除无关的甲型流感病毒抗体来制备。这是通过使抗血清通过含有来自适当毒株的破碎病毒粒子抗原的免疫吸附柱来进行的。然而，用这种方法从针对尿囊液病毒悬液的血清中获得血凝素特异性抗血清的尝试失败了。用纯化的血凝素免疫后获得的抗血清也得到了优化。这些抗血清通过免疫荧光显示在甲型流感病毒感染的亚型分型方面更具优势，但不能先验地视为亚型特异性。亚型特异性血清在临床标本上用于快速病毒诊断的实用性已得到证明。
