In Situ Point Mutation Detection in FFPE Colorectal Cancers Using the BaseScope Assay

2020 ◽  
pp. 349-360
Author(s):  
Ann-Marie Baker ◽  
Trevor A. Graham
Keyword(s):  
Point Mutation ◽  
Mutation Detection ◽  
Colorectal Cancers

scholarly journals 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 ◽  
2016 ◽  
Vol 141 (4) ◽  
pp. 1421-1433 ◽  
Author(s):  
Ceyhun E. Kirimli ◽  
Wei-Heng Shih ◽  
Wan Y. Shih
Keyword(s):  
Nucleic Acid ◽  
Point Mutation ◽  
Mutation Detection ◽  
Piezoelectric Plate ◽  
Locked Nucleic Acid ◽  
Wild Type ◽  
Single Nucleotide ◽  
Lna Probe

We have examined thein situdetection of a single-nucleotideKRASmutation 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 theKRASmutation.

Detection of gene point mutation in paraffin sections using in situ loop-mediated isothermal amplification

2007 ◽  
Vol 57 (9) ◽  
pp. 594-599 ◽  
Author(s):  
Satoshi Ikeda ◽  
Kazuhiko Takabe ◽  
Masaharu Inagaki ◽  
Naoya Funakoshi ◽  
Keiko Suzuki
Keyword(s):  
Point Mutation ◽  
Isothermal Amplification ◽  
Paraffin Sections ◽  
Loop Mediated Isothermal Amplification

scholarly journals K-ras Point Mutation Detection in Lung Cancer: Comparison of Two Approaches to Somatic Mutation Detection Using ARMS Allele-specific Amplification

2000 ◽  
Vol 46 (12) ◽  
pp. 1929-1938 ◽  
Author(s):  
Simon J Clayton ◽  
Frank M Scott ◽  
Jill Walker ◽  
Kay Callaghan ◽  
Kemal Haque ◽  
...  
Keyword(s):  
Point Mutation ◽  
Mutation Detection ◽  
Lung Tumor ◽  
Molecular Techniques ◽  
Clinical Samples ◽  
Analytical Sensitivity ◽  
Real Time Quantitative Pcr ◽  
Allele Specific ◽  
Specific Amplification ◽  
Tumor Dna

Abstract Background: The use of sensitive molecular techniques to detect rare cells in a population is of increasing interest to the molecular pathologist, but detection limits often are poorly defined in any given molecular assay. We combined the approaches of real-time quantitative PCR with ARMSTM allele-specific amplification in a novel assay for detecting mutant K-ras sequences in clinical samples. Methods: ARMS reactions were used to detect seven commonly occurring mutations in the K-ras oncogene. These mutations produce amino acid changes in codon 12 (Gly to Ala, Arg, Asp, Cys, Ser, or Val) and codon 13 (Gly to Asp). A control reaction was used to measure the total amount of amplifiable K-ras sequence in a sample so that the ratio of mutant to wild-type sequence could be measured. Quantitative data were confirmed for a selection of samples by an independent cloning and sequencing method. The assay was used to analyze 82 lung tumor DNA samples. Results: The assay detected K-ras mutations in 44% of adenocarcinomas, which is equivalent to frequencies reported in the literature using ultrasensitive techniques. Forty-six percent of squamous carcinomas were also positive. The ratio of mutant sequence in the tumor DNA samples was 0.04–100%. Conclusions: The assay is homogeneous, with addition of tumor DNA sample being the only step before results are generated. The quantitative nature of the assay can potentially be used to define the analytical sensitivity necessary for any specified diagnostic application of K-ras (or other) point mutation detection.

scholarly journals RETRACTED: Point mutation detection by economic HRM protocol primer design

2019 ◽  
Vol 18 ◽  
pp. 100628
Author(s):  
Dhafer A.F. Al-Koofee ◽  
Jawad Mohammed Ismael ◽  
Shaden M.H. Mubarak
Keyword(s):  
Point Mutation ◽  
Mutation Detection ◽  
Primer Design

Laser desorption mass spectrometry for point mutation detection

1996 ◽  
Vol 13 (4) ◽  
pp. 87-94 ◽  
Author(s):  
N.I Taranenko ◽  
K.J Matteson ◽  
C.N Chung ◽  
Y.F Zhu ◽  
L.Y Chang ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Point Mutation ◽  
Mutation Detection ◽  
Laser Desorption ◽  
Laser Desorption Mass Spectrometry ◽  
Desorption Mass Spectrometry

Eliminating the secondary structure of targeting strands for enhancement of DNA probe based low-abundance point mutation detection

2019 ◽  
Vol 1075 ◽  
pp. 137-143
Author(s):  
Zhihao Ming ◽  
Qianzhi Chen ◽  
Na Chen ◽  
Meng Lin ◽  
Na Liu ◽  
...  
Keyword(s):  
Secondary Structure ◽  
Point Mutation ◽  
Mutation Detection ◽  
Dna Probe

In-Situ Mutation Detection by Magnetic Beads-Probe Based on Single Base Extension and Its Application in Genotyping of Hepatitis B Virus Pre-C Region 1896nt Locus Single Nucleotide Polymorphisms

2019 ◽  
Vol 15 (12) ◽  
pp. 2393-2400
Author(s):  
Xianbo Mou ◽  
Danni Sheng ◽  
Zhu Chen ◽  
Mei Liu ◽  
Yuan Liu ◽  
...  
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Fluorescence Intensity ◽  
Magnetic Beads ◽  
Mutation Detection ◽  
Single Base Extension ◽  
Single Base ◽  
Base Extension ◽  
B Virus

Hepatitis B virus (HBV) is closely related to occurrence and development of viral hepatitis. A mutation of 1896nt locus in its pre-C region can promote replication of HBV DNA and improve stability of pre-genome RNA structure, and can even help HBV evade immune clearance. In this study, magnetic beads-probe (MBs@probe) method, combined with single base extension (SBE) technology, was developed for in-situ mutation detection of HBV pre-C region 1896nt locus. Before successfully completing the genotyping of 165 HBV samples, the crucial reaction conditions were first optimized, such as SBE temperature, MBs size and amount, and probe concentration on the surface of MBs. Experimental results showed that these conditions had significant effects on MBs@probe in-situ mutation detection. Comprehensive considerations, such as 58 °C of SBE temperature, high fluorescence intensity and signal-to-noise ratios (SNRs) were obtained when MBs@probe complex was made by 100 μg of 300 nm-MBs and 3.0 μM of probes in the system. Finally, 1896nt locus mutation in pre-C region of 165 HBV samples was successfully genotyped, among which 71 HBV samples were wild types and the remaining 94 samples were mutant types. Meanwhile, 14 randomly chosen samples were taken to further analyze fluorescence intensity and SNRs respectively, and sequencing results for the first two samples were consistent with results from the MBs@probe in-situ mutation detection method. Compared with two-color fluorescence hybridization (TCFH) genotyping technology, this method generally improves the SNRs to more than 10 (which is more than 2-fold), has higher reliability and is more suitable to detect SNPs for known sites.

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