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

@article{Kirimli2016AmplificationfreeIS,
  title={Amplification-free in situ KRAS point mutation detection at 60 copies per mL in urine in a background of 1000-fold wild type.},
  author={Ceyhun E. Kirimli and Wei-Heng Shih and Wan Y. Shih},
  journal={The Analyst},
  year={2016},
  volume={141 4},
  pages={
          1421-33
        }
}
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… 

Figures and Tables from this paper

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

TLDR
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.

Developing a Low-Cost, Simple-to-Use Electrochemical Sensor for the Detection of Circulating Tumour DNA in Human Fluids

TLDR
A simple, low-cost DNA biosensor that was developed specifically to detect mutations in a key oncogene (KRAS) and has the potential to be deployed in a low- cost, point-of-care test where patients can be screened either for early diagnosis purposes or monitoring of response to therapy.

Endonuclease IV based competitive DNA probe assay for differentiation of low-abundance point mutations by discriminating stable single-base mismatches.

TLDR
The unique discrimination property of Endo IV toward stable single-base mismatches located at the second nucleotide 3' to the AP site was disclosed, and a highly sensitive and specific detection system was established with discrimination factors of 510-1079 for G:X mismatches.

Installing CRISPR-Cas12a sensors in a portable glucose meter for point-of-care detection of analytes.

TLDR
The CRISPR/Cas12a-PGM system demonstrated here paves a way to further broaden the POC applications ofCRISPR-based diagnostics.

Hereditary Pancreatic Cancer

TLDR
There is a need to improve the early detection of pancreatic cancer in order to detect and improve survival in the same way that mammograms and colonoscopies have improved survival for individuals with breast and colorectal cancer.

Interested in publishing with us ? Contact book

TLDR
There is a need to improve the early detection of pancreatic cancer in order to detect and improve survival in the same way that mammograms and colonoscopies have improved survival for individuals with breast and colorectal cancer.

References

SHOWING 1-10 OF 59 REFERENCES

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.

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.

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.

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.

Locked nucleic acid (LNA) single nucleotide polymorphism (SNP) genotype analysis and validation using real-time PCR.

TLDR
Application of the three novel criteria presented provides an easy to use semi-automated genotype confirmation protocol to effectively determine ambiguous genotypes generated from a real-time PCR platform.

Transrenal DNA as a Diagnostic Tool: Important Technical Notes

TLDR
The sensitivity and specificity of detection of mutant sequences are reduced in the presence of high excess of a respective wild‐type allele, but they can be significantly increased through application of enriched polymerase chain reaction (PCR), peptide nucleic acid (PNA) clamped PCR, and/or stencil‐aided mutation analysis (SAMA), based on selective pre‐PCR elimination of wild‐ type sequences.

Single nucleotide polymorphism genotyping using short, fluorescently labeled locked nucleic acid (LNA) probes and fluorescence polarization detection.

TLDR
In this method, very short LNA probes are labeled with either rhodamine or hexachlorofluorescein, and their hybridization to target DNAs is followed by measuring the fluorescence polarization (FP) of the dyes, and the generation of a universal set of genotyping reagents is possible.
...