Logo image
Back
A locked nucleic acid clamp-mediated PCR assay for detection of a p53 codon 249 hotspot mutation in urine
Journal article   Open access   Peer reviewed

A locked nucleic acid clamp-mediated PCR assay for detection of a p53 codon 249 hotspot mutation in urine

Selena Y Lin, Veerpal Dhillon, Surbhi Jain, Ting-Tsung Chang, Chi-Tan Hu, Yih-Jyh Lin, Shun-Hua Chen, Kung-Chao Chang, Wei Song, Lixin Yu, …
The Journal of molecular diagnostics : JMD, v 13(5), pp 474-484
Sep 2011
DOI: https://doi.org/10.1016/j.jmoldx.2011.05.005
PMID: 21726666
Featured in Collection :   UN Sustainable Development Goals @ Drexel
url
https://doi.org/10.1016/j.jmoldx.2011.05.005View
Published, Version of Record (VoR)Open Access (Publisher-Specific),  Open

Abstract

Adult Aged Carcinoma, Hepatocellular - genetics Carcinoma, Hepatocellular - urine Codon - genetics DNA Mutational Analysis - methods DNA Probes - metabolism Female Humans Liver Neoplasms - genetics Liver Neoplasms - urine Male Middle Aged Mutation - genetics Nucleic Acid Denaturation - genetics Oligonucleotides - metabolism Polymerase Chain Reaction - methods Reproducibility of Results Sensitivity and Specificity Tumor Suppressor Protein p53 - genetics Tumor Suppressor Protein p53 - urine
Hepatocellular carcinoma (HCC) has a 5-year survival rate of <10% because it is difficult to diagnose early. Mutations in the TP53 gene are associated with approximately 50% of human cancers. A hotspot mutation, a G:C to T:A transversion at codon 249 (249T), may be a potential DNA marker for HCC screening because of its exclusive presence in HCC and its detection in the circulation of some patients with HCC. A locked nucleic acid clamp-mediated PCR assay, followed by melting curve analysis (using the SimpleProbe), was developed to detect the TP53 249T mutation. In this assay, the locked nucleic acid clamp suppressed 10(7) copies of wild-type templates and permitted detection of 249T-mutated template, with a sensitivity of 0.1% (1:1000) of the mutant/wild-type ratio, assessed by a reconstituted standard within 2 hours. With an amplicon size of 41 bp, it detects target DNA sequences in short fragmented DNA templates. The detected mutations were validated by DNA sequencing analysis. We then tested DNA isolated from urine samples of patients with HCC for p53 mutations and identified positive TP53 mutations in 9 of 17 samples. The possibility of using this novel TP53 249T assay to develop a urine or blood test for HCC screening is discussed.

Metrics

13 Record Views
43 citations in Web of Science
43 citations in Scopus

Details

UN Sustainable Development Goals (SDGs)

This publication has contributed to the advancement of the following goals:

#3 Good Health and Well-Being

InCites Highlights

Data related to this publication, from InCites Benchmarking & Analytics tool:

Collaboration types
Domestic collaboration
International collaboration
Web of Science research areas
Pathology
Logo image