Amplification-free in situ KRAS point mutation detection at 60 copies per mL in urine in a background of 1000-fold wild type

The Analyst
Ceyhun E KirimliWan Y Shih

Abstract

We have examined the in situ detection of a single-nucleotide KRAS mutation in urine using a (Pb(Mg1/3Nb2/3)O3)0.65(PbTiO3)0.35 (PMN-PT) piezoelectric plate sensor (PEPS) coated with a 17-nucleotide (nt) locked nucleic acid (LNA) probe DNA complementary to the KRAS mutation. To enhance the in situ mutant (MT) DNA detection specificity against the wild type (WT), detection was carried out in a flow with a flow rate of 4 mL min(-1) and at 63 °C with the PEPS vertically situated at the center of the flow in which both the temperature and the flow impingement force discriminated the wild type. Under such conditions, PEPS was shown to specifically detect KRAS MT in situ with 60 copies per mL analytical sensitivity in a background of clinically-relevant 1000-fold more WT in 30 min without DNA isolation, amplification, or labeling. For validation, this detection was followed with detection in a mixture of blue MT fluorescent reporter microspheres (FRMs) (MT FRMs) that bound to only the captured MT and orange WT FRMs that bound to only the captured WT. Microscopic examinations showed that the captured blue MT FRMs still outnumbered the orange WT FRMs by a factor of 4 to 1 even though WT was 1000-fold of MT in urine. Finally, multiplexe...Continue Reading

References

Jan 26, 2002·Science·Y D ParkB T Jonker
Dec 5, 2003·Analytical Biochemistry·Luis A UgozzoliKeith Hamby
Jul 15, 2004·Annals of the New York Academy of Sciences·Ying-Hsiu SuSamuil Umansky
Sep 30, 2004·Journal of the American Chemical Society·Rahela GasparacShana O Kelley
Jun 1, 2005·Annals of Oncology : Official Journal of the European Society for Medical Oncology·V BazanUNKNOWN Gruppo Oncologico dell'Italia Meridionale (GOIM)
Sep 20, 2005·Nature Biotechnology·Gengfeng ZhengCharles M Lieber
Mar 4, 2006·Expert Review of Molecular Diagnostics·Samuil R Umansky, L David Tomei
May 4, 2006·Nucleic Acids Research·Yong YouRichard Owczarzy
Apr 5, 2007·Analytical Chemistry·Zhiqiang GaoJinming Kong
Apr 27, 2007·Angewandte Chemie·David M HammondThomas Carell
May 29, 2007·Analytical Chemistry·Haixin ChangYifu Guan
May 4, 2010·Biosensors & Bioelectronics·Guo-Jun ZhangEu-Jin Andy Lim
Oct 19, 2010·Nucleic Acids Research·Simon A ForbesP Andrew Futreal
Mar 23, 2011·Angewandte Chemie·Tetiana KurkinaKannan Balasubramanian
Jul 26, 2011·Clinical Radiology·E G Robertson, G Baxter
Jul 4, 2012·Clinical Cancer Research : an Official Journal of the American Association for Cancer Research·Yu ImamuraShuji Ogino
Nov 14, 2012·Physical Chemistry Chemical Physics : PCCP·Masato Morita, Kaito Takahashi
Jul 10, 2013·Nature Reviews. Clinical Oncology·Emily CrowleyAlberto Bardelli
Apr 24, 2014·British Journal of Cancer·S EserD Saur

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