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单径向补体结合试验在流感、呼吸道合胞病毒、腮腺炎、腺病毒3型和单纯疱疹病毒1型感染血清学诊断中的应用。

Application of the single radial complement fixation test for serodiagnosis of influenza, respiratory syncytial, mumps, adeno type 3, and herpes simplex type 1 virus infections.

作者信息

Sato S, Ochiai H, Niwayama S

机构信息

Department of Virology, Toyama Medical and Pharmaceutical University, Japan.

出版信息

J Med Virol. 1988 Apr;24(4):395-404. doi: 10.1002/jmv.1890240406.

Abstract

A stabilized modification of the single radial complement fixation (SRCF) test in gel was developed for detecting various virus antibodies. The principle of the test is based on the use of a single-stage procedure with an agarose plate containing virus antigens and antibody-coated erythrocytes, and thin plastic film coated with complement. By filling the wells in the agar plate with a 1:4 diluted heat-inactivated sera and covering the agar surface with a complement film, a zone of unlysed cells surrounded by a hemolytic area appears after incubation overnight at 4 degrees C and then for 1-2 h at 37 degrees C, depending on the antibody titers. The SRCF antibody titer is calculated numerically from the square of the diameter of the unlysed cell zone. The stability of reagents could be significantly improved using thin complement film and several stabilizers. When this test was used for serodiagnosis of influenza, respiratory syncytial (RS), mumps, adeno virus type 3 and herpes simplex type 1 virus infections (using a total of 400 sera), excellent correlations were demonstrated for antibody titers between conventional complement fixation (CF) and SRCF titers. Furthermore, the expression of antibody titer as an SRCF unit with consecutive value, produced results sensitive to fluctuations in the antibody titers. The simplicity of the procedure, stability of the reagents, and excellent correlation with the conventional CF test might make this a useful test for routine serodiagnosis and seroepidemiological survey of various virus infections.

摘要

为检测多种病毒抗体,开发了一种凝胶中单向放射状补体结合(SRCF)试验的稳定改良方法。该试验原理基于单步程序,使用含有病毒抗原和抗体包被红细胞的琼脂糖平板,以及补体包被的薄塑料膜。通过在琼脂平板孔中加入1:4稀释的热灭活血清,并在琼脂表面覆盖补体膜,于4℃孵育过夜,然后在37℃孵育1 - 2小时后,根据抗体效价会出现一个由溶血区域包围的未裂解细胞区。SRCF抗体效价通过未裂解细胞区直径的平方数值计算得出。使用薄补体膜和几种稳定剂可显著提高试剂的稳定性。当该试验用于流感、呼吸道合胞病毒(RS)、腮腺炎、3型腺病毒和1型单纯疱疹病毒感染的血清学诊断(共使用400份血清)时,传统补体结合(CF)试验和SRCF试验的抗体效价之间显示出极好的相关性。此外,以连续值的SRCF单位表示抗体效价,所得结果对抗体效价的波动敏感。该方法操作简单、试剂稳定,且与传统CF试验相关性良好,这可能使其成为各种病毒感染常规血清学诊断和血清流行病学调查的有用试验。

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