Block copolymer-oligonucleotide conjugates for genotyping on microarrays.
Polymer-oligonucleotide conjugates were synthesized from the amphiphilic block copolymer poly(tert-butylacrylamide-b-(N-acryloylmorpholine-co-N-acryloxysuccinimide)) using an original solid-phase DNA synthesis strategy. This method provided conjugates highly functionalized with oligonucleotides throughout the polymer chain. After purification, block copolymer-oligonucleotide conjugates were spotted on a multidetection microarray system developed by Apibio using a standard nanodroplet piezo inkjet spotting technique to develop the oligosorbent assay (OLISA). Two genotyping models (HLA-DQB1 and platelet glycoproteins [GPs]), which are particularly difficult to study with standard systems, were evaluated. For both models, block copolymer-oligonucleotide conjugates used as capture probes amplified the responses of in vitro diagnostic assays. The detection limit reached by using conjugates was estimated at 15 pM for a 219-bp DNA target (HLA-DQB1 model). Moreover, single nucleotide polymorphism was detected in the platelet GPs genotyping model. The use of polymer conjugates led to a significant improvement in both sensitivity and specificity of standard hybridization assays even when applied to complex biological models.
de Lambert
Bertrand
B
Unité Mixte CNRS/bioMérieux, IFR 128 Biosciences Lyon Gerland, Ecole Normale Supérieure de Lyon, 69364 Lyon Cedex 07, France.
Chaix
Carole
C
Charreyre
Marie-Thérèse
MT
Martin
Thibault
T
Aigoui
Arnaud
A
Perrin-Rubens
Agnès
A
Pichot
Christian
C
Mandrand
Bernard
B
D000180:Acrylic Resins / Q000737:chemistry*; D005838:Genotype; D006683:HLA-DQ Antigens / Q000235:genetics*; D059866:HLA-DQ beta-Chains; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation* / Q000379:methods; D009841:Oligonucleotides / Q000737:chemistry*; D010980:Platelet Membrane Glycoproteins / Q000235:genetics*; D020641:Polymorphism, Single Nucleotide; D013050:Spectrometry, Fluorescence
D000180:Acrylic Resins; D006683:HLA-DQ Antigens; D059866:HLA-DQ beta-Chains; C069051:HLA-DQB1 antigen; D009841:Oligonucleotides; D010980:Platelet Membrane Glycoproteins; C527051:poly(tert-butylacrylamide-b-(N-acryloylmorpholine-co-N-acryloxysuccinimide))