A piezoelectric affinity biosensor for genetically modified organisms (GMOs) detection

被引:23
作者
Minunni, M
Tombelli, S
Pratesi, S
Mascini, M
Piatti, P
Bogani, P
Buiatti, M
Mascini, M
机构
[1] Univ Florence, Dipartimento Sanita Pubbl Epidemiol & Chim Anal A, I-50121 Florence, Italy
[2] Univ Teramo, Fac Agr, I-64023 Mosciano Stazione, Italy
[3] Lab Chim Camera Commercio Torino, I-10127 Turin, Italy
[4] Univ Florence, Dipartinento Biol Anim & Genet, I-50127 Florence, Italy
关键词
piezoelectric biosensors; DNA; hybridisation; genetically modified organisms; soy;
D O I
10.1081/AL-100103595
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
A piezoelectric affinity sensor, based on DNA hybridisation has been studied for applications to Genetically Modified Organisms (GMOs) detection. The thiol/dextran modified surfaces were coupled to streptavidin for immobilising 5'-biotinyltead probes (25-mer). The probes sequences were respectively internal to the amplified product of P35S and T-NOS. These target sequences were chosen on the base of their wide presence in GMOs. The system has been optimised using synthetic complementary oligonucleotides (25-mer) and the specificity of the system tested with a noncomplementary oligonucleotide (23-mer). The hybridisation study was performed also with samples of DNA isolated from CRM (Certified Reference Materials) soybean powder containing 2% of transgenic material and amplified by PCR. Non amplified genomic or plasmidic DNA was also used. The developed system was very specific, binding only the complementary DNA strand. The CV% was 20% both with synthetic oligonucleotides and PCR amplified samples. The sensor signal was independent of the sample dilution but the system is still at a semi-quantitative level.
引用
收藏
页码:825 / 840
页数:16
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