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使用补体片的单向放射补体结合试验。流感病毒株和型特异性抗原抗体反应的测定。

Single radial complement fixation test using complement film. Assay of the antibody response to strain and type specific antigens of influenza virus.

作者信息

Sato S, Motoda S, Iwase I, Jo K

出版信息

J Virol Methods. 1983 Aug;7(2):57-64. doi: 10.1016/0166-0934(83)90092-7.

Abstract

A stabilized modification of the single radial complement fixation test in gel (SRCF) was developed for detecting influenza antibodies. The principle of the test is the use of a single-step procedure with the following reagents: (1) Agarose plate containing influenza antigen and antibody coated erythrocytes (EA). (2) Thin plastic film coated with dried complement. By filling the wells cut in the agar with the heat inactivated serum samples and covering the agar surface with the complement film, a zone of unlysed cells surrounded by a haemolytic area appears after overnight incubation for 16-18 h at 4 degrees C and 1-2 h at 37 degrees C. The squares of the zone diameter were measured for estimating the antibody quantity by using CF(S) and virion antigen of influenza virus, and the type-specific antibody was demonstrated by using CF(S) antigen, while the strain-specific antibody was demonstrated by using virion antigen. An excellent correlation was demonstrated for antibody titres between conventional CF and SRCF with CF(S) antigen, on the one hand, and, between conventional HI and SRCF with virion antigen, on the other.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

为检测流感抗体,开发了一种凝胶中单向放射补体结合试验(SRCF)的稳定改良方法。该试验原理是采用单步程序,使用以下试剂:(1)含有流感抗原和抗体包被红细胞(EA)的琼脂糖平板。(2)涂有干燥补体的薄塑料膜。通过在琼脂上切出的孔中加入热灭活血清样本,并用补体膜覆盖琼脂表面,在4℃过夜孵育16 - 18小时,然后在37℃孵育1 - 2小时后,会出现一个未裂解细胞区,周围是溶血区。测量该区域直径的平方,通过使用流感病毒的CF(S)和病毒粒子抗原估计抗体量,使用CF(S)抗原可证明型特异性抗体,使用病毒粒子抗原可证明株特异性抗体。一方面,传统补体结合试验(CF)与使用CF(S)抗原的SRCF之间的抗体滴度,另一方面,传统血凝抑制试验(HI)与使用病毒粒子抗原的SRCF之间的抗体滴度,均显示出极好的相关性。(摘要截短至250字)

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