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  • 标题:Development of a high-throughput cell based 384-well influenza A quantification assay for interpandemic and highly pathogenic avian strains
  • 本地全文:下载
  • 作者:Melicia R. Gainey ; Ann M. Wasko ; Jennifer N. Garver
  • 期刊名称:Health
  • 印刷版ISSN:1949-4998
  • 电子版ISSN:1949-5005
  • 出版年度:2010
  • 卷号:2
  • 期号:1
  • 页码:32-37
  • DOI:10.4236/health.2010.21006
  • 出版社:Scientific Research Publishing
  • 摘要:Influenza remains a world wide health threat, thus the need for a high-throughput and robust assay to quantify both seasonal and avian in-fluenza A strains. Therefore, a 384-well plate format was developed for the median tissue culture infectious dose assay (TCID50) utilizing the detection of nucleoprotein by an in situ en-zyme linked immunosorbent assay (ELISA) which was optimized for sensitivity in this assay. Highly pathogenic avian influenza, A/Vietnam/ 1203/04 (H5N1), and interpandemic strains, A/ New Caledonia/20/99 (H1N1) and A/Brisbane/ 10/07 (H3N2), were quantified using this high- throughput assay. Each 384-well plate can be used to analyze ten viral samples in quadrupli-cate, eight dilutions per sample, including all necessary assay controls. The results obtained from 384-well plates were comparable to tradi-tional 96-well plates and also demonstrate re-peatability, intermediate precision, and assay linearity. Further, the use of 384-well plates in-creased the throughput of sample analysis and the precision and accuracy of the resulting titer.
  • 关键词:Avian Influenza; ELISA; High-Throughput Assay; Interpandemic Influenza A; TCID50
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