Simulated Respiratory Secretion for Use in the Development of Influenza Diagnostic Assays
Simulated Respiratory Secretion for Use in the Development of Influenza Diagnostic Assays
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DOI：
10.1371/journal.pone.0166800
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发表时间：
2016-11-21
期刊：
影响因子：
3.7
通讯作者：
Henrickson, Kelly J.
中科院分区：
文献类型：
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作者：
Bose, Michael E.;McCaul, Kate C.;Henrickson, Kelly J.
流感病毒是一种重要的呼吸道病原体，目前已有许多检测方法。这些检测方法的发展通常涉及使用人类临床样本来验证其性能。然而，临床样品可能难以获得，随着时间的推移而恶化，并且成分不一致。本研究的目的是开发一种模拟呼吸分泌物（SRS），可以作为临床样品的替代品。为此，我们确定了主要的呼吸分泌成分（Na+、K+、Ca2+、细胞、白蛋白、IgG、IgM和粘蛋白）对流感检测（包括核酸扩增和快速抗原检测）性能的影响。除血清源性人IgG抑制快速抗原检测信号外，所有检测组分对分子检测的影响都很小。使用点印迹法，我们能够显示抗流感核蛋白IgG抗体在人类呼吸道样本中是常见的。我们制作了一个含有人体呼吸样本成分中点水平的SRS，并在Veritor、Sofia、CDC RT-PCR、Simplexa、cobas Liat和Alere i流感检测中研究了其与磷酸盐缓冲盐水和病毒阴性临床样本基质的效果。我们的研究结果表明，SRS可以以类似于临床样品的方式与各种测试方法相互作用，对测试性能产生类似的影响。
Many assays have been developed for the detection of influenza virus which is an important respiratory pathogen. Development of these assays commonly involves the use of human clinical samples for validation of their performance. However, clinical samples can be difficult to obtain, deteriorate over time, and be inconsistent in composition. The goal of this study was to develop a simulated respiratory secretion (SRS) that could act as a surrogate for clinical samples. To this end, we determined the effects major respiratory secretion components (Na+, K+, Ca2+, cells, albumin IgG, IgM, and mucin) have on the performance of influenza assays including both nucleic acid amplification and rapid antigen assays. Minimal effects on the molecular assays were observed for all of the components tested, except for serum derived human IgG, which suppressed the signal of the rapid antigen assays. Using dot blots we were able to show anti-influenza nucleoprotein IgG antibodies are common in human respiratory samples. We composed a SRS that contained mid-point levels of human respiratory sample components and studied its effect compared to phosphate buffered saline and virus negative clinical sample matrix on the Veritor, Sofia, CDC RT-PCR, Simplexa, cobas Liat, and Alere i influenza assays. Our results demonstrated that a SRS can interact with a variety of test methods in a similar manner to clinical samples with a similar impact on test performance.