Scientific Affairs, Luminex, A DiaSorin Company, 12212 Technology Blvd., Austin, TX 78727, USA.
Methods. 2023 Nov;219:22-29. doi: 10.1016/j.ymeth.2023.09.003. Epub 2023 Sep 15.
Multiplexing in biological assays allows the simultaneous detection of multiple analytes in a single reaction, which reduces time, labor, and cost as compared to single reaction-based detection methods. Microsphere- or bead-based suspension array technologies, such as the Luminex® xMAP® system, offer high-throughput detection of nucleic acids through a variety of different assay chemistries. Common with most nucleic acid chemistries, for bead-based or other microarray technologies, is the need for efficient extraction and purification of the nucleic acids from the specimen of interest. Often, the optimal method will be dictated by the requirements of the up-front enzymatic chemistry, such as PCR, primer extension, branched DNA (bDNA), etc. For bead-based microarray platforms, the user must also be cognizant of proteins and other contaminants present in reactions that require heat denaturation, as that can lead to bead aggregation or agglutination, preventing the reading of assay results. This review describes and highlights some of the nucleic acid extraction and purification methods that have been used successfully for bead-based nucleic acid analysis, for both prokaryotic and eucaryotic nucleic acids, from a variety of sample types.
多重分析在生物检测中允许在单个反应中同时检测多个分析物，与基于单反应的检测方法相比，可减少时间、劳动力和成本。基于微球或珠的悬浮阵列技术，如 Luminex® xMAP®系统，通过多种不同的检测化学方法提供核酸的高通量检测。对于基于珠或其他微阵列技术的大多数核酸化学方法而言，常见的是需要从感兴趣的标本中有效提取和纯化核酸。通常，最佳方法将取决于酶化学的前期要求，例如 PCR、引物延伸、分支 DNA（bDNA）等。对于基于珠的微阵列平台，用户还必须意识到在需要热变性的反应中存在的蛋白质和其他污染物，因为这可能导致珠聚集或凝集，从而阻止检测结果的读取。本综述描述并强调了一些已成功用于基于珠的核酸分析的核酸提取和纯化方法，这些方法适用于来自各种样本类型的原核和真核核酸。
