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Sources of variance in suspended microarray assays and Intraplex method for improving precision and repeatability.

机译：悬浮微阵列测定法和Intraplex方法的方差来源可提高精度和可重复性。

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摘要

Flow cytometry based suspended microarray assays are susceptible to error in the assays and instrument. I present the Intraplexing method for compensating for these sources of error, evidence for the major sources of variance, and straightforward models which suggest other minor contributing factors of error. Intraplexes require little sample, are inexpensive, sensitive, and quantify within statistical confidence limits.;The Intraplex method uses ratios of sensitivity in multiplexed assay sets to the same analyte within a sample. The intraplex method compensates for the sources of variance that have been identified in suspended microarray assays. It requires no changes to instruments to construct precision assays.;The major sources of error are: The suspended microarray system generates statistical distributions of counts. The number of microbeads picked up for different bead regions (sets) will vary in a classic bell curve. This problem can be ignored with small numbers of multiplexed assays, but it is significant when the number of assays multiplexed together rises.;The microparticles on which assays are conducted vary significantly in size.;Random carryover of microbeads between wells is an intractable source of error. I show that instruments carry over microbeads in sufficient numbers to be considered a statistically valid sample. This source of error is the probable primary reason why it has been so difficult for suspended microarray assays to get through approval as diagnostics.;Opto-electronics may not respond similarly on different instruments. The opto-electronic system does not necessarily respond identically between different instruments at differing signal intensity. Inter-instrument ratio can be unity for calibration point and two logs for lower intensity signal.;There are other sources of error. Aside from microsphere agglutination, they appear to be minor contributors compensated for by the Intraplex method; however, these sources of error deserve further study.;Intraplexing can enable reliable, precise, time course studies in small animals for virtually any assay of interest. If Intraplexing is used, then the assay should be suitable for use in human diagnostics, as well as making for better science. I believe the Intraplexing method should become standard practice for assay development and analysis.
机译：基于流式细胞仪的悬浮微阵列测定法易受测定法和仪器误差的影响。我介绍了用于补偿这些误差源的内部复用方法，主要方差证据以及表明其他次要误差因素的简单模型。 Intraplex几乎不需要样品，价格便宜，灵敏并且可以在统计置信范围内进行定量。Intraplex方法使用了多重化验集中灵敏度与样品中相同分析物的比率。 intraplex方法补偿了在悬浮微阵列分析中已经确定的差异来源。它不需要改变用于构建精密测定的仪器。主要的误差来源是：悬浮微阵列系统生成计数的统计分布。为不同的珠子区域（组）采集的微珠的数量将按照经典的钟形曲线变化。这个问题可以通过少量的多重分析来忽略，但是当多重分析的数目增加时，这个问题就显得很重要。进行分析的微粒的大小差异很大。错误。我表明仪器携带的微珠数量足以被视为统计上有效的样品。这种错误的来源可能是导致悬浮微阵列分析难以通过诊断的重要原因。光电在不同仪器上的响应可能不同。光电系统不一定会在不同仪器之间以不同的信号强度做出相同的响应。仪器间的比率对于校准点可以是统一的，对于较低强度的信号可以是两个对数。；还有其他误差源。除微球凝集外，它们似乎是通过Intraplex方法补偿的较小贡献者。但是，这些错误来源值得进一步研究。内部复杂化可以在小型动物中进行可靠，精确的时程研究，从而几乎可以进行任何感兴趣的测定。如果使用Intraplexing，则该测定应适合用于人类诊断以及更好的科学。我相信内混法应该成为测定开发和分析的标准方法。

著录项

  • 作者

    Hanley, Brian Paul.;

  • 作者单位

    University of California, Davis.;

  • 授予单位 University of California, Davis.;
  • 学科 Biology Biostatistics.;Biology Microbiology.;Biophysics Medical.
  • 学位 Ph.D.
  • 年度 2009
  • 页码 47 p.
  • 总页数 47
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

  • 入库时间 2022-08-17 11:38:25

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