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Coupling of a DNA piezoelectric biosensor and polymerase chain reaction to detect apolipoprotein E polymorphisms
University Florence, Dipartimento Sanita Pubbl Epidemiol and Chim Analit, Florence, Italy; INRCA, Lab Biol Mol, Florence, Italy; Cranfield University, Cranfield Biotechnol Centre, Cranfield MK43 0AL, Beds, England; .
University Florence, Dipartimento Sanita Pubbl Epidemiol and Chim Analit, Florence, Italy; INRCA, Lab Biol Mol, Florence, Italy; Cranfield University, Cranfield Biotechnol Centre, Cranfield MK43 0AL, Beds, England; .
University Florence, Dipartimento Sanita Pubbl Epidemiol and Chim Analit, Florence, Italy; INRCA, Lab Biol Mol, Florence, Italy; Cranfield University, Cranfield Biotechnol Centre, Cranfield MK43 0AL, Beds, England; .
University Florence, Dipartimento Sanita Pubbl Epidemiol and Chim Analit, Florence, Italy; INRCA, Lab Biol Mol, Florence, Italy; Cranfield University, Cranfield Biotechnol Centre, Cranfield MK43 0AL, Beds, England; .
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2000 (English)In: Biosensors & bioelectronics, ISSN 0956-5663, E-ISSN 1873-4235, Vol. 15, no 08-jul, 363-370 p.Article in journal (Refereed) Published
Abstract [en]

In this paper we report the coupling of the Polymerase Chain Reaction (PCR) with a piezoelectric biosensor to detect a point mutation in a human gene. Biotinylated 23-mer probes were immobilised on the streptavidin coated gold surface of a quartz crystal; streptavidin was covalently bound to the thiol/dextran modified gold surface. The hybridisation of the immobilised probes with a short sequence (23 mer) complementary, non-complementary and mismatched DNA was investigated: the device was able to distinguish the different synthetic oligonucleotides. Many cycles of measurements can be performed on the same crystal surface regenerating the single strand of DNA with 1 mM of HCl. The same hybridisation reaction was then performed using real samples of human DNA extracted from blood and amplified by PCR, following a standard procedure for genetic detection of the polymorphism of the apolipoprotein E (apoE) gene. The procedure was able to distinguish the sequences present in the different samples, which differ only in one base: in this way it was possible distinguish between different groups of genotypes with apoE typing. Experiments with blank samples confirmed the absence of adsorption or non-specific effects on the quartz crystal treated with the reported procedure. (C) 2000 Elsevier Science S.A. All rights reserved.

Place, publisher, year, edition, pages
Elsevier Science B.V., Amsterdam. , 2000. Vol. 15, no 08-jul, 363-370 p.
Keyword [en]
piezoelectric biosensor; PCR; hybridisation; polymorphism; apolipoprotein E
National Category
Engineering and Technology
Identifiers
URN: urn:nbn:se:liu:diva-65231ISI: 000165079200005OAI: oai:DiVA.org:liu-65231DiVA: diva2:395003
Available from: 2011-02-04 Created: 2011-02-04 Last updated: 2017-12-11

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