Piezoelectric Plate Sensor (PEPS) for Analysis of Specific KRAS Point Mutations at Low Copy Number in Urine Without DNA Isolation or Amplification.

@article{Kirimli2017PiezoelectricPS,
  title={Piezoelectric Plate Sensor (PEPS) for Analysis of Specific KRAS Point Mutations at Low Copy Number in Urine Without DNA Isolation or Amplification.},
  author={Ceyhun E. Kirimli and Wei-Heng Shih and Wan Y. Shih},
  journal={Methods in molecular biology},
  year={2017},
  volume={1572},
  pages={
          327-348
        }
}
We have examined in situ detection of single-nucleotide KRAS mutations 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 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 °C with the PEPS vertically situated at the center of the flow. Both… 
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References

SHOWING 1-10 OF 40 REFERENCES
Amplification-free in situ KRAS point mutation detection at 60 copies per mL in urine in a background of 1000-fold wild type.
TLDR
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.
Specific in situ hepatitis B viral double mutation (HBVDM) detection in urine with 60 copies ml(-1) analytical sensitivity in a background of 250-fold wild type without DNA isolation and amplification.
TLDR
In situ detection of hepatitis B virus 1762T/1764A double mutation (HBVDM) in urine using a (PMN-PT) piezoelectric plate sensor coated with a 16-nucleotide (nt) probe DNA (pDNA) complementary to the HBVDM is examined.
Temperature- and flow-enhanced detection specificity of mutated DNA against the wild type with reporter microspheres.
TLDR
It was shown that at room temperature, flow initially increased the binding of both the MT and WT at lower flow rates but decreased the binding at flow rates ≥4 ml min(-1) due to the increase in the flow-induced impingement force on the FRMs to overcome the bound pDNA and the WT to the GCG at higher flow rates.
DNA hybridization detection with 100 zM sensitivity using piezoelectric plate sensors with an improved noise-reduction algorithm.
TLDR
Real-time, in situ hybridization detection of target DNA (tDNA) in a buffer solution and in urine using 8 μm-thick lead magnesium niobate-lead titanate (PMN-PT) piezoelectric plate sensors (PEPSs) and a new multiple-parabola (>50) resonance peak position fitting algorithm is examined.
PEMC-based method of measuring DNA hybridization at femtomolar concentration directly in human serum and in the presence of copious noncomplementary strands.
Piezoelectric-excited, millimeter-sized cantilever (PEMC) sensors having high-mode resonance near 1 MHz are shown to exhibit mass change sensitivity of 1-300 ag/Hz. Gold-coated PEMC sensors
Detection of the factor V Leiden mutation by direct allele-specific hybridization of PCR amplicons to photoimmobilized locked nucleic acids.
TLDR
The simplicity of the assay and its procedural relatedness to the widely used ELISA format should make it useful for routine factor V testing in the clinical laboratory.
Piezoelectric plate sensor for in situ genetic detection of hepatitis b virus in serum without DNA isolation and amplification
TLDR
A new type of sensor developed in Shih’s laboratory that is unique is that the shift is caused by the stress generated by the binding of the target and is inherently amplified by the polarization switching in the PMN-PT layer.
DNA melting analysis for detection of single nucleotide polymorphisms.
TLDR
DNA melting analysis (DM) was used successfully for variant detection, and two previously unknown SNPs were discovered by this approach, making it highly suitable for the large-scale detection of sequence variants.
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