Strömberg Mattias, Zardán Gómez de la Torre Teresa, Göransson Jenny, Gunnarsson Klas, Nilsson Mats, Svedlindh Peter, Strømme Maria
Department of Engineering Sciences, Division of Nanotechnology and Functional Materials, Uppsala University, The Angstrom Laboratory, Box 534, SE-751 21 Uppsala, Sweden.
Anal Chem. 2009 May 1;81(9):3398-406. doi: 10.1021/ac900561r.
The possibility for conducting multiplex detection of DNA-sequences using the volume-amplified magnetic nanobead detection assay [Stromberg, M.; Goransson, J.; Gunnarsson, K.; Nilsson, M.; Svedlindh, P. and Strømme, M. Nano Lett. 2008 , 8, 816-821] was investigated. In this methodology, a batch consisting of a mixture of several sizes of probe-tagged magnetic beads was used for detection of several types of targets in the same compartment. Furthermore, a nonlinear least-squares deconvolution procedure of the composite imaginary part of complex magnetization vs frequency spectra based on the Cole-Cole model was applied to analyze the data. The results of a quantitative biplex analysis experiment were compared with the corresponding separate single-target assays. Finally, triplex analysis was briefly demonstrated qualitatively. Biplex and triplex detection were found to perform well qualitatively. Biplex detection was found to enable a rough target quantification. Multiplex detection may become a complement to performing multiple separate single-target assays for, e.g., parallel detection of multiple infectious pathogens. Multiplex detection also permits robust relative quantification and inclusion of an internal control to improve quantification accuracy.
研究了使用体积放大磁纳米珠检测法[斯特伦伯格，M.；戈兰松，J.；贡纳松，K.；尼尔松，M.；斯韦德林德，P.和斯特勒姆，M.《纳米快报》2008年，8卷，816 - 821页]进行DNA序列多重检测的可能性。在这种方法中，由几种大小的探针标记磁珠混合物组成的一批用于在同一隔室中检测几种类型的靶标。此外，基于科尔 - 科尔模型对复磁化强度的复虚部与频率谱进行非线性最小二乘反褶积程序来分析数据。将定量双靶标分析实验的结果与相应的单独单靶标检测进行比较。最后，定性地简要展示了三靶标分析。发现双靶标和三靶标检测在定性方面表现良好。发现双靶标检测能够进行大致的靶标定量。多重检测可能成为对例如多种传染性病原体的平行检测进行多个单独单靶标检测的一种补充。多重检测还允许进行可靠的相对定量并纳入内部对照以提高定量准确性。
