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血清中寄生虫特异性人IgG和IgE的定量:固相放射免疫分析和酶联免疫吸附测定方法的评估

The quantitation of parasite-specific human IgG and IgE in sera: evaluation of solid-phase RIA and ELISA methodology.

作者信息

Hamilton R G, Hussain R, Ottesen E A, Adkinson N F

出版信息

J Immunol Methods. 1981;44(1):101-14. doi: 10.1016/0022-1759(81)90111-3.

Abstract

We have developed a non-competitive solid-phase radioimmunoassay (SPRIA) to quantitate both human IgE and IgG antibodies against soluble adult antigens of Brugia malayi (B.m.), a filarial parasite causing extensive infection throughout the tropics. Previously enzyme-linked immunosorbent assays (ELISA) had been used to detect microgram/ml levels of IgG anti-B.m., but IgE antibodies were difficult to detect in this system. Since the SPRIA successfully quantitates both IgG and IgE anti-B.m., we sought to examine the reasons for the SPRIA's apparent superiority in detecting IgE anti-B.m. by extracting specific IgG from sera with high levels of IgE and IgG anti-B.m. antibodies. IgE anti-B.m. was then quantitated in these sera using both the SPRIA and ELISA methods. Results indicate that IgG anti-B.m. does not interfere with detection of specific IgE antibody in the SPRIA but does interfere in the ELISA. While ELISA permits detection of IgE anti-B.m. in the absence of competing IgG anti-B.m., as levels of specific IgG increase, the IgE is no longer detectable. These differences between SPRIA and ELISA can be explained by the SPRIA's antigen excess conditions which assure that there are sufficient antigens both to detect all anti-B.m. antibodies present in the serum and to adequately represent all antigen specificities in the crude B.m. extract. Our findings commend the use of SPRIA methods over ELISA in assessment of B.m.-specific IgE antibody in filariasis and indicate a potential role for SPRIA methods in absolute quantitation of specific serum antibodies.

摘要

我们开发了一种非竞争性固相放射免疫分析方法(SPRIA),用于定量检测人体针对马来布鲁线虫(B.m.)可溶性成虫抗原的IgE和IgG抗体。马来布鲁线虫是一种丝虫寄生虫,在整个热带地区引发广泛感染。此前,酶联免疫吸附测定法(ELISA)已被用于检测微克/毫升水平的抗B.m. IgG,但在该系统中难以检测到IgE抗体。由于SPRIA能够成功定量抗B.m. IgG和IgE,我们试图通过从含有高水平抗B.m. IgE和IgG抗体的血清中提取特异性IgG,来探究SPRIA在检测抗B.m. IgE方面明显更具优势的原因。然后使用SPRIA和ELISA方法对这些血清中的抗B.m. IgE进行定量。结果表明,抗B.m. IgG在SPRIA中不干扰特异性IgE抗体的检测,但在ELISA中会产生干扰。虽然ELISA在不存在竞争性抗B.m. IgG的情况下能够检测到抗B.m. IgE,但随着特异性IgG水平的升高,IgE就不再能被检测到。SPRIA和ELISA之间的这些差异可以通过SPRIA的抗原过量条件来解释,该条件可确保有足够的抗原既能检测血清中存在的所有抗B.m.抗体,又能充分代表粗制B.m.提取物中的所有抗原特异性。我们的研究结果表明,在评估丝虫病中抗B.m.特异性IgE抗体时,SPRIA方法优于ELISA方法,并表明SPRIA方法在绝对定量特异性血清抗体方面具有潜在作用。

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