title: Simultaneous detection of multiple mutations in epidermal growth factor receptor based on fluorescence quenching of quantum dots.
abstract: We have developed a simultaneous detection method for two common mutations in the epidermal growth factor receptor gene based on the fluorescence quenching phenomenon caused by aggregation of CdSe quantum dots. For detection of the in-frame deletion in exon 19 and the L858R point mutation in exon 21, water-soluble CdSe quantum dots with two sizes were functionalized using four different types of probe oligonucleotides. Addition of target oligonucleotides with the deletion mutation in exon 19 into the suspensions caused crosslinking-induced aggregation of green-emitting quantum dots, followed by the fluorescence quenching while that with the L858R point mutation resulted in aggregation of yellow-emitting quantum dots. In addition, targets with both deletion and point mutations caused aggregation of both green- and yellow-emitting quantum dots. This method allows a simultaneous detection of mutations in exon 19 and 21 of EGFR gene in a single experiment. We found that minimum mutant concentration that could be detected by this method was as low as 2% for deletion mutation, and 5% for point mutation. PCR products of EGFR gene were also used to confirm that our method could be used to detect mutation in amplified DNA fragments.
authors.affiliation: Department of Chemical and Biomolecular Engineering, Yonsei University, Seoul, Republic of Korea.
authors.forename: Taegyeong
authors.initials: T
authors.lastname: Kang
authors.forename: Hosub
authors.initials: H
authors.lastname: Lee
authors.forename: Dongcheol
authors.initials: D
authors.lastname: Choe
authors.forename: Sang-Woo
authors.initials: SW
authors.lastname: Joo
authors.forename: So Yeong
authors.initials: SY
authors.lastname: Lee
authors.forename: Kyong-Ah
authors.initials: KA
authors.lastname: Yoon
authors.forename: Kangtaek
authors.initials: K
authors.lastname: Lee
mesh_terms: D004252:DNA Mutational Analysis / Q000295:instrumentation*; D015342:DNA Probes / Q000235:genetics*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D066246:ErbB Receptors / Q000235:genetics*; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation*; D045663:Quantum Dots; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence / Q000295:instrumentation*
chemical_list: D015342:DNA Probes; D066246:ErbB Receptors