Use of bovine viral diarrhoea virus as an internal control for amplification of hepatitis C virus
Use of bovine viral diarrhoea virus as an internal control for amplification of hepatitis C virus
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DOI：
10.1159/000031044
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发表时间：
1999-01-01
期刊：
影响因子：
2.7
通讯作者：
Simmonds, P
中科院分区：
文献类型：
--
作者：
Cleland, A;Nettleton, P;Simmonds, P
背景和目的：在欧洲和其他地区，通过聚合酶链式反应(PCR)筛查丙型肝炎病毒(HCV)将成为筛查献血者和血浆献血者的强制性要求。这项研究描述了一种内部控制的、高灵敏度的聚合酶链式反应方法，旨在筛查池中的献血者。材料和方法：以从牛病毒性腹泻病毒(BVDV)中提取的RNA为内对照，监测丙型肝炎病毒聚合酶链式反应中提取、逆转录和扩增步骤的效率。结果：在存在33个BVDV RNA的基因组等价物的情况下，通过降低BVDV的扩增效率，获得了对单分子丙型肝炎病毒的敏感性。用超速离心法和NucliSens提取法可从大容量池(2-5ml)中高效地回收BVDV。结论：BVDV的检测验证了提取、逆转录和扩增法检测血浆中丙型肝炎病毒的有效性，为阴性结果的检测提供了有价值的质量保证。
Background and Objectives: Screening for hepatitis C virus (HCV) by polymerase chain reaction (PCR) will be mandatory for screening blood and plasma donors in Europe and elsewhere. This study describes an internally controlled, highly sensitive PCR method designed for screening blood donations in pools. Material and Methods: RNA extracted from bovine viral diarrhoea virus (BVDV) was used as an internal control to monitor the efficiency of extraction, reverse transcription and amplification steps in HCV PCR. Results: Sensitivity of PCR for single molecules of HCV in the presence of 33 genome equivalents of BVDV RNA was achieved by reducing the efficiency of BVDV amplification. BVDV could be recovered at high efficiency from large volume pools (2-5 ml) by ultracentrifugation and by the NucliSens extraction method. Conclusion: Detection of BVDV validates the extraction, reverse transcription and amplification methods used for HCV detection in plasma pools and provides valuable quality assurance for negative results.