Saldanha J, Minor P
Department of Virology, National Institute for Biological Standards and Control, Hertfordshire, England.
J Med Virol. 1994 May;43(1):72-6. doi: 10.1002/jmv.1890430114.
Although current manufacturing processes appear to efficiently inactivate hepatitis C virus (HCV), it is possible that contaminated blood products may result from failure of some stage during manufacture or from virus overload of plasma pools used for preparation of products. While antibody screening probably removes the majority of HCV positive donations, some donations which are antibody-negative but HCV positive may be included in pools. The HCV RNA content of plasma pools from paid and voluntary donors was investigated by polymerase chain reaction (PCR). A sensitive PCR method using a single pair of primers from the 5' non-coding region of the HCV genome and a "hot-start" was established and shown to be as sensitive as the more conventional nested PCR (which uses two pairs of primers). The majority of pools from paid donors (prescreening) were HCV RNA positive, while all pools from voluntary donors were both antibody and RNA negative. Intravenous immunoglobulins prepared from contaminated pools were RNA negative despite having high antibody levels, indicating satisfactory clearance of the virus during manufacture. The virus load of the pools was at least a thousand-fold lower than that of single donations, possibly as a result of treatment during the production of the pools or the presence of factors in pools which reduce the sensitivity of some part of the PCR assay. The HCV content of a plasma donation was determined as 3.6 x 10(6) genomes/ml by an end point dilution method. Thus a simple and sensitive PCR assay was established for detecting HCV RNA in plasma pools and blood products.
尽管目前的生产工艺似乎能有效地灭活丙型肝炎病毒（HCV），但在生产过程中，某些环节的失误或用于制品制备的血浆池病毒过载仍有可能导致血液制品被污染。虽然抗体筛查可能会筛除大部分HCV阳性献血，但一些抗体阴性但HCV阳性的献血可能会被混入血浆池中。采用聚合酶链反应（PCR）对有偿献血者和自愿献血者血浆池中的HCV RNA含量进行了研究。建立了一种使用来自HCV基因组5'非编码区的一对引物及“热启动”的灵敏PCR方法，结果表明该方法与更传统的巢式PCR（使用两对引物）一样灵敏。大部分有偿献血者（预检）的血浆池HCV RNA呈阳性，而所有自愿献血者的血浆池抗体和RNA均为阴性。尽管由受污染血浆池制备的静脉注射免疫球蛋白抗体水平很高，但其RNA为阴性，这表明在生产过程中病毒得到了满意的清除。血浆池的病毒载量至少比单次献血低一千倍，这可能是由于血浆池生产过程中的处理或血浆池中存在降低PCR检测某些部分灵敏度的因素所致。采用终点稀释法测定一份血浆捐献的HCV含量为3.6×10(6)个基因组/ml。因此，建立了一种简单灵敏的PCR检测方法用于检测血浆池和血液制品中的HCV RNA。
