Stolz M, Gilgen M, Niederhauser C
Blood Transfusion Service SRC Berne, Switzerland.
Vox Sang. 2003 Feb;84(2):105-10. doi: 10.1046/j.1423-0410.2003.00263.x.
Hepatitis C virus-polymerase chain reaction (HCV-PCR) minipool testing can improve the safety of labile blood products owing to a reduction in the diagnostic preseroconversion window period. In Switzerland, HCV-PCR minipool testing for the release of labile blood components became mandatory in September 1999. In the largest Swiss blood transfusion centre, HCV-PCR minipool testing began in January 1999. This report analyses the performance of the test during a 3-year period: 1 January 1999 to 31 December 2001.
EDTA-blood was collected in either standard tubes or plasma preparation (PPT) tubes from 10 blood transfusion services in Switzerland and then sent to the Blood Transfusion Service SRC Berne. Up to 48 donor samples were pooled overnight using Tecan Genesis RSP 200/8 pipettors. Viral RNA was extracted by using the Qiagen QIAamp 96 viral RNA BioRobot kit on a BioRobot 9604. For PCR amplification and detection of HCV or internal control (IC) sequences, the Roche Cobas Amplicor v2.0 test kit was used. Data management, pool resolution and identification of positive samples were performed using the PMS Software from Tecan.
In the 3-year period from 1 January 1999 to 31 December 2001, 839056 blood donor samples were tested in minipools of up to 48 samples. Thirty-five HCV-PCR-positive donations were identified. Thirty-four samples had antibodies against HCV and were therefore also detected by screening for antibody to HCV (anti-HCV). In October 2001, one seronegative (but PCR-positive) donor was detected.
HCV-PCR minipool testing was successfully introduced in the largest Swiss blood transfusion service. It was shown that the release of HCV-PCR minipool results can be accomplished concurrently with the results of serological analysis. The challenge with a seronegative, but PCR-positive, donor demonstrates that the minipool testing strategy adds additional safety to blood products.
丙型肝炎病毒聚合酶链反应（HCV-PCR）混合样本检测可缩短诊断窗口期，从而提高不稳定血液制品的安全性。1999年9月起，瑞士强制要求对不稳定血液成分进行HCV-PCR混合样本检测。在瑞士最大的输血中心，HCV-PCR混合样本检测于1999年1月开始。本报告分析了1999年1月1日至2001年12月31日这3年期间该检测的性能。
从瑞士10个输血服务机构采集的乙二胺四乙酸（EDTA）抗凝血样本，分装于标准管或血浆制备（PPT）管中，然后送至伯尔尼输血服务中心SRC。使用Tecan Genesis RSP 200/8移液器将多达48份供者样本混合过夜。采用Qiagen QIAamp 96病毒RNA自动提取试剂盒在BioRobot 9604仪器上提取病毒RNA。使用罗氏Cobas Amplicor v2.0检测试剂盒进行HCV或内对照（IC）序列的PCR扩增和检测。使用Tecan公司的PMS软件进行数据管理、混合样本拆分及阳性样本鉴定。
在1999年1月1日至2001年12月31日的3年期间，共对839056份供者血液样本进行了混合样本检测，每份混合样本最多包含48份样本。共鉴定出35份HCV-PCR阳性捐献样本。其中34份样本抗HCV抗体呈阳性，因此通过抗HCV抗体筛查也可检测到。2001年10月，检测到1名血清学阴性（但PCR阳性）的供者。
瑞士最大的输血服务机构成功引入了HCV-PCR混合样本检测。结果表明，HCV-PCR混合样本检测结果可与血清学分析结果同时得出。血清学阴性但PCR阳性的供者样本带来的挑战表明，混合样本检测策略为血液制品增添了额外的安全性。
