NanoHDA : A nanoparticle-assisted isothermal amplification technique for genotyping assays

Research output: Journal Publications and Reviews (RGC: 21, 22, 62)21_Publication in refereed journalpeer-review

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Author(s)

Detail(s)

Original languageEnglish
Pages (from-to)12-21
Journal / PublicationNano Research
Volume10
Issue number1
Publication statusPublished - 1 Jan 2017
Externally publishedYes

Abstract

Isothermal methods, such as helicase-dependent amplification (HDA), have an advantage over polymerase chain reaction for DNA amplification owing to their ease of operation. Here, we developed a new HDA method that is nanoparticle-assisted, termed nanoHDA. This method uses gold nanoparticles (AuNPs) to improve the sensitivity and specificity of the isothermal method. In HDA, the denaturation of DNA templates is mediated by helicases, but this method is limited by the low denaturation efficiency of helicases. In this report, AuNPs with preferential affinity for single-stranded DNA (ssDNA) were utilized to improve the denaturation efficiency of helicases. The same affinity property of nanoparticles can also enhance specificity by suppressing primer-dimer formation. This nanoHDA method was employed to genotype the KRAS gene in genomic DNA samples from colorectal cancer patients, as achieved by the hybridization of nanoHDA amplicons using the NanoBioArray chip. [Figure not available: see fulltext.]

Research Area(s)

  • colorectal cancer, genotyping assay, gold nanoparticle, helicase, isothermal DNA amplification, KRAS, single nucleotide polymorphism

Bibliographic Note

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Citation Format(s)

NanoHDA : A nanoparticle-assisted isothermal amplification technique for genotyping assays. / Sedighi, Abootaleb; Oberc, Christopher; Whitehall, Vicki; Li, Paul C. H.

In: Nano Research, Vol. 10, No. 1, 01.01.2017, p. 12-21.

Research output: Journal Publications and Reviews (RGC: 21, 22, 62)21_Publication in refereed journalpeer-review