Multiplex Detection of DNA Mutations by the Fluorescence Fingerprint Spectrum Technique

摘要

Detection of DNA mutations is clinically important for diagnostics, prognostics, and disease assessment. A variety of methods have been developed for the detection of DNA mutations, such as DNA sequencing (dideoxy sequencing and pyrosequencing), high-resolution melting analysis (HRMA), and qRT-PCR methods. However, these methods are only suitable for mutation detection on a single gene or exon, which cannot achieve multiplexed analysis easily. As the demands of clinical diagnostics, prognostics, and disease assessment increase, it is appealing to develop methods that allow simultaneous detection of multiple genetic alterations. Single-base primer extension assay (such as ABI PRISM SNaPshot Multiplex Kit, Applied Biosystems) is such a highly attractive diagnostic method. However, it needs expensive capillary sequencer as well as sophisticated post-extension treatment. SNaPshot genotyping is suitable for single base substitution detection, but it is not applicable for the detection of complex mutations (such as deletion and insertion mutation). MALDI-TOF (matrix-assisted laser desorption/ionization time-of-flight) mass spectrometry can offer high multiplexing capability for DNA mutation detection; however, it requires expensive instruments and technical expertise. Therefore, it is highly desirable to develop new method for simple, rapid, sensitive, and flexible detection of multiple DNA mutations.