Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
23
pubmed:dateCreated
2008-12-8
pubmed:abstractText
An unsurpassed level of sensitivity was reached in the detection of minority mutated alleles. A low-density microarray was printed on a substrate specifically designed to provide an interference effect which amplifies the collection of the light emitted on the support and reinforces the intensity of excitation light. Optimal performance of the array was obtained by maximizing the probe density and the binding efficiency to the target through a polymeric coating made by the adsorption of a copolymer of N,N-dimethylacrylamide (97% of moles), N,N-acryloyloxysuccinimide (2%) and 3-(trimethoxysilyl)propyl methacrylate (1%) synthesized by free radical copolymerization. The new substrate was used in the identification of fetal mutations in the maternal plasma DNA. Amino-modified amplicons from genomic DNA corresponding to the locus of eight beta-thalassemia mutations were immobilized and interrogated with dual-color oligonucleotide targets. Compared with the conventional glass substrates, the new substrate showed a great enhancement of fluorescence signals thanks to the combination of the optics with the highly efficient polymeric coating, allowing specific detection of all mutations. The high sensitivity and selectivity obtained made it possible to develop assays for the identification of paternally inherited mutations on fetal DNA in the maternal plasma in couples at risk for beta-thalassemia.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Dec
pubmed:issn
0173-0835
pubmed:author
pubmed:issnType
Print
pubmed:volume
29
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
4714-22
pubmed:meshHeading
pubmed-meshheading:19053069-Alleles, pubmed-meshheading:19053069-Base Sequence, pubmed-meshheading:19053069-DNA, pubmed-meshheading:19053069-DNA Mutational Analysis, pubmed-meshheading:19053069-DNA Primers, pubmed-meshheading:19053069-Electrophoresis, Microchip, pubmed-meshheading:19053069-Female, pubmed-meshheading:19053069-Fetal Diseases, pubmed-meshheading:19053069-Genotype, pubmed-meshheading:19053069-Humans, pubmed-meshheading:19053069-Male, pubmed-meshheading:19053069-Mutation, pubmed-meshheading:19053069-Oligonucleotide Array Sequence Analysis, pubmed-meshheading:19053069-Paternity, pubmed-meshheading:19053069-Polymerase Chain Reaction, pubmed-meshheading:19053069-Polymers, pubmed-meshheading:19053069-Pregnancy, pubmed-meshheading:19053069-Prenatal Diagnosis, pubmed-meshheading:19053069-beta-Globins, pubmed-meshheading:19053069-beta-Thalassemia
pubmed:year
2008
pubmed:articleTitle
Development of new substrates for high-sensitive genotyping of minority mutated alleles.
pubmed:affiliation
Genomic Unit for the Diagnosis of Human Pathologies, San Raffaele Scientific Institute, Milan, Italy.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't