Oligonucleotide ligation assay-based DNA chip for multiplex detection of single nucleotide polymorphism.
Biosens Bioelectron
; 19(10): 1277-83, 2004 May 15.
Article
em En
| MEDLINE
| ID: mdl-15046760
ABSTRACT
An oligonucleotide ligation assay-based DNA chip has been developed to detect single nucleotide polymorphism. Synthesized nonamers, complementary to the flanking sequences of the mutation sites in target DNA, were immobilized onto glass slides through disulfide bonds on their 5' terminus. Allele-specific pentamers annealed adjacent to the nonamers on the complementary target DNA, containing 5'-phosphate groups and biotin labeled 3'-ends, were mixed with the target DNA in tube. Ligation reactions between nonamers and pentamers were carried out on chips in the presence of T4 DNA ligase. Ligation products were directly visualized on chips through enzyme-linked assay. The effect of GT mismatch at different positions of pentamers on the ligation were evaluated. The results showed that any mismatch between pentamer and the target DNA could lead to the decrease of ligation, which can be detected easily. The established approach was further used for multiplex detection of mutations in rpoB gene of rifampin-resistant Mycobacterium tuberculosis clinical isolates.
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Base de dados:
MEDLINE
Assunto principal:
DNA
/
Análise de Sequência com Séries de Oligonucleotídeos
/
Polimorfismo de Nucleotídeo Único
Tipo de estudo:
Diagnostic_studies
Idioma:
En
Ano de publicação:
2004
Tipo de documento:
Article