Saldanha J
N.I.B.S.C., Division of Virology, Potters Bar, Herts, UK.
Dev Biol Stand. 1993;81:231-6.
Plasma pools and blood products were analysed for hepatitis C virus (HCV) RNA by the polymerase chain reaction (PCR). Initially, samples were amplified by "nested" PCR using two pairs of primers from the highly-conserved 5' non-coding region of the genome. The method was later modified to amplification with only the outer pair of primers and a "hot-start" to eliminate amplification of non-target sequences. The modified method was as sensitive as "nested" PCR. However, with samples containing low levels of RNA, it is necessary to perform repeat amplifications to avoid false-negative results. Ten out of 17 anti-HCV positive pools from paid donors were positive, while none of the 12% pools from unpaid donors was positive for RNA or anti-HCV antibodies. Four i.v. immunoglobulins made from contaminated pools were virus-free, indicating removal of virus during current manufacturing processes. A quantitative PCR method was established using a positive donation. Probit analysis gave an estimated viral titre of 3.6 x 10(6) genomes/ml.
采用聚合酶链反应（PCR）对血浆池和血液制品进行丙型肝炎病毒（HCV）RNA分析。最初，使用来自基因组高度保守的5'非编码区的两对引物，通过“巢式”PCR对样本进行扩增。该方法后来改进为仅使用外侧引物对进行扩增，并采用“热启动”以消除非靶序列的扩增。改进后的方法与“巢式”PCR一样灵敏。然而，对于RNA含量低的样本，有必要进行重复扩增以避免假阴性结果。来自有偿献血者的17个抗-HCV阳性血浆池中，有10个呈阳性，而来自无偿献血者的12%血浆池中，没有一个RNA或抗-HCV抗体呈阳性。由受污染血浆池制成的4种静脉注射免疫球蛋白无病毒，表明在当前生产过程中病毒已被去除。使用一份阳性献血样本建立了定量PCR方法。概率分析得出估计病毒滴度为3.6×10⁶基因组/毫升。
