Wunschel D S, Fox K F, Fox A, Bruce J E, Muddiman D C, Smith R D
Department of Microbiology and Immunology, University of South Carolina, School of Medicine, Columbia 29209, USA.
Rapid Commun Mass Spectrom. 1996;10(1):29-35. doi: 10.1002/(SICI)1097-0231(19960115)10:1<29::AID-RCM430>3.0.CO;2-#.
The analysis of polymerase chain reaction (PCR) products by electrospray ionization-Fourier transform ion cyclotron resonance mass spectrometry (ESI-FTICR) has been achieved. Specifically, a 105 base-pair nucleotide portion of the ribosomal spacer region was amplified in two members of the B. cereus group (i.e. B. thuringiensis and B. cereus) using PCR. These amplified regions were then analyzed by gel electrophoresis and ESI-FTICR. Based on the predicted sequence of the PCR products for each organism, the mass measurement using ESI-FTICR matched the theoretical mass within experimental error and was consistent with gel electrophoresis results. In contrast, for the typical several hour time-scale of the gel electrophoresis experiment, the mass spectrometric analysis was completed in a matter of minutes. To our knowledge, this constitutes the first report demonstrating the ionization and detection of a double-stranded PCR product by ESI-MS. This preliminary result indicates the potential use of ESI-MS to analyze PCR products on a rapid time-scale, with potential for medical and taxonomic applications.
已实现通过电喷雾电离 - 傅里叶变换离子回旋共振质谱（ESI - FTICR）对聚合酶链反应（PCR）产物进行分析。具体而言，使用PCR在蜡样芽孢杆菌组的两个成员（即苏云金芽孢杆菌和蜡样芽孢杆菌）中扩增核糖体间隔区的105个碱基对的核苷酸部分。然后通过凝胶电泳和ESI - FTICR对这些扩增区域进行分析。基于每个生物体PCR产物的预测序列，使用ESI - FTICR进行的质量测量在实验误差范围内与理论质量相符，并且与凝胶电泳结果一致。相比之下，对于凝胶电泳实验通常需要数小时的时间尺度，质谱分析在几分钟内即可完成。据我们所知，这是第一份证明通过ESI - MS对双链PCR产物进行电离和检测的报告。这一初步结果表明ESI - MS在快速时间尺度上分析PCR产物具有潜在用途，在医学和分类学应用方面具有潜力。
