Novel Application of SELDI-TOF Mass Spectrometer in Genotyping and Personalized Medicine
M Jin, S Yang, H Wu. Ohio State University, Columbus, OH
Background: SNPs are genetic markers that determine an individual's susceptibility to various diseases. SNPs have also been used as molecular markers to predict drug efficacy and safety. We postulate that SELDI-TOF mass spectrometer offers an excellent platform for genotyping medically important SNPs for diagnosis and for prediction of therapeutic responses. This hypothesis was tested in several clinical settings, including 1) SNPs influencing warfarin drug sensitivity and safety, 2) SNPs (CYP2D6) predicting patient response to tamoxifen; 3) SNPs (CYP2C19) impacting Plavix's drug efficacy, and 4) SNPs determining genetic risks for clotting disorders.
Design: DNA containing each target SNP was first amplified by PCR using specific primers. Afterwards, single base extension was performed to generate specific SNP product. Finally, genetic variants displaying different masses are bound to Q10 anionic proteinChips and then genotyped using a SELDI-TOF mass spectrometer in a multiplexed fashion.
Results: SELDI-TOF mass spectrometer offers unique properties of on-chip sample enrichment and clean-ups, which streamline the testing procedures and eliminate many tedious experimental steps required by the conventional mass spectrometer based method. The turnaround time by SELDI-TOF mass spectrometer, from sample collection to the report of genotypes for each panel of SNPs, is less than five hours. The analytical accuracy of this method has been confirmed both by bidirectional DNA sequencing and by comparing the genotype results obtained by SELDI-TOF mass spectrometer to reports from a clinical reference laboratory.
Conclusions: By using SELDI-TOF mass spectrometer, we have devised a novel multiplex genotyping method that is fast, accurate, cost effective, and will improve patient care and clinical outcomes through personalized health care.
Monday, March 22, 2010 9:30 AM
Poster Session I Stowell-Orbison/Surgical Pathology/Autopsy Awards Poster Session # 255, Monday Morning