DEVELOPMENT OF SCREENING TESTS FOR HTLV-III

本研究旨在开发基于流式细胞仪的 用于检测特异性IgG的免疫反应性珠（IRB）测定， IgM抗体对rDNA产生的HIV抗原。 均匀聚苯乙烯珠 用四种测定抗原（p31、gp 120、p24和gp 41）包被， 每种抗原使用不同大小的珠粒。 所述涂覆的珠粒是 与稀释的血清样品和结合的免疫球蛋白一起孵育 用异硫氰酸荧光素-F（ab '）2抗人IgG鉴定 或IgM。 然后用FACSCAN（Becton-2000）分析染色的珠粒。 Dickenson）根据珠粒大小。 具有不同荧光素的珠粒 也可以以20，000个珠的速率分离和计数缀合物 在不到20秒内。 以蛋白质印迹法为标准， IRB测定显示100%灵敏度和100%特异性。 此外，62例酶免疫测定（EIA）假阳性中有61例，WB- 阴性标本经IRB检测为阴性。 18例EIA阳性的WB- 可疑血清中，14份被鉴定为HIV阴性，1份被鉴定为HIV阳性 通过IRB测定。 因此，该测定是灵敏的、特异的、快速的， 适应自动化。 最重要的是，检测艾滋病毒感染 可以更早地通过诸如基于IgM检测的测定来进行 而不是通过其他目前可用的测定。 制定一项 自动血液筛查系统（ABSS）在血液中的实际应用 中心.

This research addresses the development of flow cytometer-based immunoreactive bead (IRB) assay for the detection of specific IgG and IgM antibodies to rDNA-produced HIV antigens. Uniform polystyrene beads are coated with four assay antigens (p31, gp120, p24 and gp41), with a different sized bead used for each antigen. The coated beads are incubated with diluted serum samples and the bound immunoglobulin identified by using fluorescein isothiocyanate-F (ab')2 anti-human IgG or IgM. The stained beads are then analyzed with FACSCAN (Becton- Dickenson) according to bead size. Beads with different fluorescein conjugates can also be separated and counted at a rate of 20,000 beads in less than 20 seconds. Using the Western blot assay as the standard, the IRB assay exhibited 100% sensitivity and 100% specificity. Furthermore, 61 of the 62 enzyme immunoassay (EIA) false positive, WB- negative specimens were negative by IRB assay. Of 18 EIA-positive WB- equivocal sera, 14 were identified as HIV-negative and 1 as HIV-positive by IRB assay. Thus, the assay is sensitive, specific, rapid, and adaptable to automation. Most importantly, detection of HIV infection may be made much earlier by an assay such as this based on IgM detection than by other currently available assays. The development of an automated blood screening system (ABSS) practically useful in blood centers.