DETECTION AND CHARACTERIZATION OF HCV RNA SEQUENCES IN IMMUNE GLOBULINS

免疫球蛋白中 HCV RNA 序列的检测和表征

• 批准号: 3748300
• 负责人: M W YU
• 金额: --
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3748300
• 关键词: antiviral antibody; blood fractionation; blood preservation; drug quality /standard; drug screening /evaluation; hepatitis C virus; human tissue; immunoglobulin G; immunoglobulins; nucleic acid sequence; polymerase chain reaction; virus RNA; virus genetics

In February 1994, Baxter Healthcare Corporation, withdrew all of their Immune Globulin Intravenous (Human) (IGIV) lots which include Gammaguard (manufactured from commercial plasma, both Source Plasma and recovered plasma) and Polygam (recovered plasma from American Red cross) because of the reports of possible hepatitis C virus (HCV) transmission. Since then, we have been working intensively with Baxter's IGIV and sera from some recipients, and tried to understand the rationale for such outbreak. Previously we did not detect HCV RNA in any of the 3 lots of Gammaguard and 2 lots of Polygam made during 1988-89. Thus far, we have assayed a total of 40 lots of Gammaguard by PCR. Reconstituted samples from 31 lots were also given to Hepatitis Laboratory (HL) for PCR and for assaying anti-HCV reactivity by RIBA-2. Gammaguard lots manufactured since 1992 were found to be anti-HCV negative. For lots manufactured in 1993, we detected HCV RNA in 15 of 26 lots; among them, 21 lots were assayed by both labs, and we detected 12 positive while HL detected 13 positive (PCR results from 14 lots were agreeable between 2 labs). For lots made during 1988-1992, we found HCV RNA only in 1 of 14 lots; among 10 lots assayed by both labs, we found 1 positive while HL found the same lot and additonal 2 lots positive. All of our 15 positive lots and 12 of HL's 13 positive lots were manufactured since April 1993, which coincides with the time when Baxter manufactured IGIV lots 100% derived from multiantigen anti-HCV screened plasma. HCV RNA was also detected in 16 of 18 recipients'sera. Therefore, we strongly suspect that protective antibodies for HCV might have been removed from IGIV because of multiantigen screening of plasma. We found 1 of 4 gammaguard lots prepared from recovered plasma was positive for HCV RNA while HL found 3 lots positive. However, by limiting dilution we identified 8 lots prepared from Source Plasma having higher levels of HCV RNA. We are in the process of characterizing the HCV RNA present in 2 implicated lots which we determined to have the highest levels of HCV RNA among all.

1994 年 2 月，百特医疗保健公司撤回了所有 免疫球蛋白静脉注射（人）（IGIV）批次，其中包括 Gammaguard （由商业血浆制成，包括源血浆和回收血浆 血浆）和 Polygam（从美国红十字会回收的血浆），因为 可能传播丙型肝炎病毒（HCV）的报告。 自从 然后，我们一直在与 Baxter 的 IGIV 和血清进行深入合作 一些接受者，并试图了解这种爆发的原因。 此前我们在 3 个批次的 Gammaguard 中均未检测到 HCV RNA 以及 1988-89 年间制作的 2 批宝丽金 (Polygam)。 到目前为止，我们已经测试了一个 通过 PCR 检测总共 40 批 Gammaguard。 重构样品来自 31 个 批次也被送到肝炎实验室 (HL) 进行 PCR 和 通过 RIBA-2 测定抗 HCV 反应性。 Gammaguard 批量生产 自1992年以来，检测结果为抗HCV阴性。 对于制造批次 1993年，我们在26个批次中的15个批次中检测到了HCV RNA；其中，21个地块 两个实验室均进行了检测，我们检测到 12 例呈阳性，而 HL 检测到 13 例 阳性（2 个实验室 14 个批次的 PCR 结果一致）。 为了 在 1988-1992 年间生产的批次中，我们仅在 14 批次中的 1 批次中发现了 HCV RNA；之中 两个实验室检测了 10 个批次，我们发现 1 个呈阳性，而 HL 也发现了相同的情况 批次和额外 2 批次呈阳性。 我们所有 15 个阳性批次和 12 个 HL 的 13 个阳性批次中有 1 个是自 1993 年 4 月以来生产的，其中 与 Baxter 生产 100% 衍生 IGIV 批次的时间一致 来自多抗原抗 HCV 筛选血浆。 还检测到HCV RNA 18 名接受者中的 16 名接受者的血清中。 因此，我们强烈怀疑 HCV 保护性抗体可能已从 IGIV 中删除，因为 血浆多抗原筛查。 我们找到了 4 个伽马防护批次中的 1 个 从回收的血浆中制备的 HCV RNA 呈阳性，而 HL 发现 3批次呈阳性。 然而，通过有限稀释，我们确定了 8 个批次 由具有较高水平的 HCV RNA 的源血浆制备。我们在 2 个相关批次中 HCV RNA 的表征过程 我们确定其 HCV RNA 水平最高。