PARTITIONING OF HEPETITIS C VIRUS DURING COHN-ONCLEY FRACTION OF PLASMA

丙型肝炎病毒在血浆 COHN-ONCLEY 部分中的分离

• 批准号: 3792650
• 负责人: M W YU
• 金额: --
• 依托单位: CENTER BIOLOGICS EVALUATION RESEARCH HEMATOLOGY
• 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/3792650
• 关键词: blood donor; blood fractionation; communicable disease transmission; hepatitis C virus; human tissue; immunoglobulin G; mass screening; plasma; polymerase chain reaction; precipitation; virus RNA

Because of concern about the safety of immunoglobulins with respect to transmission of hepatitis C, we examined the partitioning of hepatitis C virus (HCV) during alcohol fractionation of a plasma pool prepared exclusively from anti-HCV reactive donations. Quantitation of HCV RNA was accomplished by nested polymerase chain reaction (PCR) at limiting dilution. One PCR unit was arbitrarily defined as the minimum amount of HCV RNA from which an amplified product could be detected. The sensitivity of the PCR assay for HCV RNA was determined by performing limiting dilution analysis on a sample of infectious plasma (H strain) known to contain 106.6 chimpanzee infectious doses/ml. In our assay, this sample contained 1.4 x 106 PCR units of HCV RNA/ml. Thus, the PCR assay has a sensitivity comparable to the chimpanzee model. When 3,073 plasma donations from otherwise acceptable donors were tested for anti-HCV, 186 were repeatedly reactive and 2,887 were negative. A pool prepared from the anti-HCV reactive donations contained 1.4 x 105 PCR units of HCV RNA/ml, whereas a pool prepared from the negative donations contained 1.6 x 103 PCR units/ml. It can be calculated that a pool comprised of all 3,073 units would contain 1.0 x 104 PCR units/ ml. Thus, in this instance, anti-HCV screening decreased the viral load of the plasma pool by a factor of 6. A 100-ml sample of the anti-HCV reactive pool was fractionated to immune globulin by the Cohn-Oncley procedure, and samples of the various fractions were analyzed for HCV RNA. Most of the HCV RNA was found in cryoprecipitate and in Cohn fractions I and III, but it was also detected in fraction II, used for immunoglobulin G preparations. A 3.4% solution of IgG prepared from this fraction II contained 30 PCR units/ml. The fractionation process leading to immune globulin resulted in an overall reduction in HCV RNA by a factor of 4.7 x 104. Although the presence of HCV RNA in the final product does not necessarily imply the presence of virus, this work suggests that the safety of immune globulins with respect to HCV transmission is not due solely to partitioning of HCV away from the immunoglobulin fraction.

由于担心免疫球蛋白的安全性， 丙型肝炎的传播，我们检查了丙型肝炎的分区 制备的血浆池乙醇分馏期间的HCV病毒（HCV） 仅来自抗HCV反应性捐赠。 HCV RNA的定量是 通过限制性巢式聚合酶链反应（PCR）完成 稀释 一个PCR单位被任意定义为最小量的 可检测到扩增产物的HCV RNA。 灵敏度 通过进行有限稀释， 对已知含有106.6的感染性血浆（H株）样品的分析 黑猩猩感染剂量/毫升 在我们的测定中，该样品含有1.4 x 106 PCR单位HCV RNA/ml。 因此，PCR测定具有灵敏度 与黑猩猩模型相似。 当3，073份血浆捐赠来自 对其他可接受的供体进行抗HCV检测，186例重复检测 2,887人为阴性。 由抗HCV抗体制备的合并液 反应性捐献物含有1.4 × 105 PCR单位的HCV RNA/ml，而 从阴性供体制备的合并液含有1.6 × 103 PCR单位/ml。 可以计算出，由所有3，073个单元组成的池将包含 1.0 x 104个PCR单位/ ml。 因此，在这种情况下，抗HCV筛查 使血浆池的病毒载量降低6倍。 的100-ml 抗-HCV反应池的样品通过以下方法分级分离成免疫球蛋白： Cohn-Oncley程序，并将各种级分的样品 分析HCV RNA。 大部分HCV RNA存在于冷沉淀中， 在Cohn级分I和III中，但也在级分II中检测到， 免疫球蛋白G的制备。 制备3.4% IgG溶液， 该级分II含有30个PCR单位/ml。 分馏工艺 导致免疫球蛋白导致HCV RNA总体减少， 系数为4.7 × 104。 尽管HCV RNA在最终的 产品并不一定意味着病毒的存在，这项工作 提示免疫球蛋白对HCV的安全性 HCV的传播不仅仅是由于HCV从细胞中分离出来， 免疫球蛋白部分。