LigAmp for sensitive detection of single-nucleotide differences.
Publication/Presentation Date
11-1-2004
Abstract
We developed the LigAmp assay for sensitive detection and accurate quantification of viruses and cells with single-base mutations. In LigAmp, two oligonucleotides are hybridized adjacently to a DNA template. One oligonucleotide matches the target sequence and contains a probe sequence. If the target sequence is present, the oligonucleotides are ligated together and detected using real-time PCR. LigAmp detected KRAS2 mutant DNA at 0.01% in mixtures of different cell lines. KRAS2 mutations were also detected in pancreatic duct juice from patients with pancreatic cancer. LigAmp detected the K103N HIV-1 drug resistance mutation at 0.01% in plasmid mixtures and at approximately 0.1% in DNA amplified from plasma HIV-1. Detection in both systems is linear over a broad dynamic range. Preliminary evidence indicates that reactions can be multiplexed. This assay may find applications in the diagnosis of genetic disorders and the management of patients with cancer and infectious diseases.
Volume
1
Issue
2
First Page
141
Last Page
147
ISSN
1548-7091
Published In/Presented At
Shi, C., Eshleman, S. H., Jones, D., Fukushima, N., Hua, L., Parker, A. R., Yeo, C. J., Hruban, R. H., Goggins, M. G., & Eshleman, J. R. (2004). LigAmp for sensitive detection of single-nucleotide differences. Nature methods, 1(2), 141–147. https://doi.org/10.1038/nmeth713
Disciplines
Business Administration, Management, and Operations | Health and Medical Administration | Management Sciences and Quantitative Methods
PubMedID
15782177
Department(s)
Administration and Leadership
Document Type
Article