K-ras mutation detection by hybridization to a polypyrrole DNA chip.
Clin Chem
; 47(2): 186-94, 2001 Feb.
Article
en En
| MEDLINE
| ID: mdl-11159765
ABSTRACT
BACKGROUND:
Detection of mutations in cancer-related genes is of major importance for both basic knowledge and clinical practice. Several strategies have been developed to diagnose these alterations. We describe a method based on polypyrrole DNA chip technology to detect K-ras gene mutations in tumors.METHODS:
An oligodeoxynucleotide array was constructed on a silicon device by copolymerization of 5'-pyrrole-labeled oligodeoxynucleotides and pyrrole. The samples to be analyzed were then amplified by PCR, and the single-stranded biotin-labeled amplified DNA was specifically hybridized to the addressed probes. Perfectly matched duplexes were detected by fluorescence microscopy using R-phycoerythrin as the detection label. The developed methodology was applied to genotype assignment of K-ras in human samples. The genotypes of 75 DNA genomic samples from colorectal cancer patients were analyzed side by side using direct DNA sequencing and a polypyrrole DNA chip.RESULTS:
The chip method unequivocally defined all of the genotypes. Mutations present at <10% of the wild-type DNA concentration could be distinguished.CONCLUSIONS:
This probe array assay is a rapid and reliable procedure that may be used to detect mutations.
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Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Polímeros
/
Pirroles
/
ADN de Neoplasias
/
Neoplasias Colorrectales
/
Proteína Oncogénica p21(ras)
/
Genes ras
/
Análisis de Secuencia por Matrices de Oligonucleótidos
Tipo de estudio:
Diagnostic_studies
Límite:
Humans
Idioma:
En
Revista:
Clin Chem
Asunto de la revista:
QUIMICA CLINICA
Año:
2001
Tipo del documento:
Article
País de afiliación:
Francia