Book chapter
Piezoelectric Plate Sensor (PEPS) for Analysis of Specific KRAS Point Mutations at Low Copy Number in Urine Without DNA Isolation or Amplification
Biosensors and Biodetection, pp 327-348
01 Jan 2017
PMID: 28299698
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
We have examined in situ detection of single-nucleotide KRAS mutations in urine using a (Pb(Mg1/3Nb2/3)O-3)(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 without DNA isolation and amplification. In situ mutant (MT) DNA in urine in a wild type (WT) background was carried out at a flow rate of 4 mL/min and at 63 degrees C with the PEPS vertically situated at the center of the flow. Both the temperature and the impingement flow force discriminated the wild type. Under these conditions PEPS was shown to specifically detect KRAS MT in situ within 30 min with an analytical sensitivity of 60 copies/mL in a clinically relevant background of WT with concentrations 1000-fold greater than that of MT without DNA isolation, amplification, or labeling. For validation, detection was performed in a mixture of blue MT fluorescent reporter microspheres (FRMs) (MT FRMs) that bound to only the captured MT, and orange WTFRMs that bound to only the captured WT. The captured blue MTFRMs still outnumbered the orange WT FRMs by a factor of 4-1 even though WT was 1000-fold of MT in urine, illustrating the specificity of the point mutation detection.
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Details
- Title
- Piezoelectric Plate Sensor (PEPS) for Analysis of Specific KRAS Point Mutations at Low Copy Number in Urine Without DNA Isolation or Amplification
- Creators
- Ceyhun E. Kirimli - Drexel UniversityWei-Heng Shih - Drexel UniversityWan Y. Shih - Drexel University
- Contributors
- B Prickril (Editor)A Rasooly (Editor)
- Publication Details
- Biosensors and Biodetection, pp 327-348
- Series
- Methods in Molecular Biology
- Publisher
- Springer Nature; DORDRECHT
- Number of pages
- 22
- Grant note
- R41 AI112224; R41 AI120445 / NIAID NIH HHS; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Institute of Allergy & Infectious Diseases (NIAID)
- Resource Type
- Book chapter
- Language
- English
- Academic Unit
- School of Biomedical Engineering, Science, and Health Systems; Materials Science and Engineering
- Web of Science ID
- WOS:000444594800023
- Scopus ID
- 2-s2.0-85015679700
- Other Identifier
- 991019167475904721
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InCites Highlights
Data related to this publication, from InCites Benchmarking & Analytics tool:
- Web of Science research areas
- Biochemical Research Methods
- Biochemistry & Molecular Biology