title: Kras gene codon 12 mutation detection enabled by gold nanoparticles conducted in a nanobioarray chip.
abstract: This study employs a nanobioarray (NBA) chip for multiple biodetection of single base pair mutations at the Kras gene codon 12. To distinguish between the mutant and wild-type target DNAs, current bioarray methods use high-temperature hybridization of the targets to the allele-specific probes. However, these techniques need prior temperature optimization and become harder to implement in the case of the detection of multiple mutations. We aimed to detect these mutations at a single temperature (room temperature), enabled by the use of gold nanoparticles (AuNPs) on the bioarray created within nanofluidic channels. In this method, a low amount of target oligonucleotides (5fmol) and polymerase chain reaction (PCR) products (300pg) were first loaded on the AuNP surface, and then these AuNP-bound targets were introduced into the channels of a polydimethylsiloxane (PDMS) glass chip. The targets hybridized to their complementary probes at the intersection of the target channels to the pre-printed oligonucleotide probe lines on the glass surface, creating a bioarray. Using this technique, fast and high-throughput multiple discrimination of the Kras gene codon 12 were achieved at room temperature using the NBA chip, and the specificity of the method was proved to be as high as that with the temperature stringency method.
authors.affiliation: Department of Chemistry, Simon Fraser University, Burnaby, British Columbia V5A 1S6, Canada.
authors.forename: Abootaleb
authors.initials: A
authors.lastname: Sedighi
authors.affiliation: Department of Chemistry, Simon Fraser University, Burnaby, British Columbia V5A 1S6, Canada. Electronic address: paulli@sfu.ca.
authors.forename: Paul C H
authors.initials: PC
authors.lastname: Li
mesh_terms: D000483:Alleles; D003062:Codon; D004247:DNA / Q000032:analysis*; D005821:Genetic Techniques / Q000295:instrumentation*; D006046:Gold / Q000737:chemistry*; D006801:Humans; D053768:Metal Nanoparticles / Q000737:chemistry*; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide; D011518:Proto-Oncogene Proteins / Q000235:genetics* / Q000378:metabolism; D016283:Proto-Oncogene Proteins p21(ras); D013696:Temperature; D018631:ras Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D003062:Codon; C117307:KRAS protein, human; D011518:Proto-Oncogene Proteins; D006046:Gold; D004247:DNA; D016283:Proto-Oncogene Proteins p21(ras); D018631:ras Proteins
keywords: Gold nanoparticle; Kras gene; Microarray; Nanobioarray chip; Single nucleotide polymorphism; Target DNA