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Magnetic bead technology for viral RNA extraction from serum in blood bank screening

机译:磁珠技术在血库筛选中从血清中提取病毒RNA

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摘要

Nucleic acid amplification testing (NAT) was recently recommended by Brazilian legislation and has been implemented at some blood banks in the city of São Paulo, Brazil, in an attempt to reduce blood-born transmission of human immunodeficiency virus (HIV) and hepatitis C virus. OBJECTIVE: Manual magnetic particle-based extraction methods for HIV and HCV viral nucleic acids were evaluated in combination with detection by reverse transcriptase - polymerase chain reaction (RT-PCR) one-step. METHODS: Blood donor samples were collected from January 2010 to September 2010, and minipools of them were submitted to testing. ELISA was used for the analysis of anti-HCV/HIV antibodies. Detection and amplification of viral RNA was performed using real-time PCR. RESULTS: Out of 20.808 samples screened, 53 samples (29 for HCV and 24 for HIV) were confirmed as positive by serological and NAT methods. CONCLUSION: The manual magnetic bead-based extraction in combination with real-time PCR detection can be used to routinely screen blood donation for viremic donors to further increase the safety of blood products.
机译:巴西立法最近推荐了核酸扩增测试(NAT),并已在巴西圣保罗市的一些血库中实施,以减少人类免疫缺陷病毒(HIV)和丙型肝炎病毒的血源性传播。目的:通过一步一步结合逆转录酶-聚合酶链反应(RT-PCR)检测,评估了基于手动磁性粒子的HIV和HCV病毒核酸提取方法。方法:从2010年1月至2010年9月收集献血者样本,并将其中的小样本池进行测试。 ELISA用于分析抗HCV / HIV抗体。使用实时PCR进行病毒RNA的检测和扩增。结果:在筛选的20.808个样本中,通过血清学和NAT方法证实了53个样本(HCV为29个,HIV为24个)为阳性。结论:基于手动磁珠的提取结合实时PCR检测可用于常规筛查病毒血症供体的献血,以进一步提高血液制品的安全性。

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