A DNA piezoelectric biosensor assay coupled with a polymerase chain reaction for bacterial toxicity determination in environmental samples
2000 (English)In: Analytica Chimica Acta, ISSN 0003-2670, E-ISSN 1873-4324, Vol. 418, no 1Article in journal (Refereed) Published
In this paper, we report the realisation of a DNA piezoelectric biosensor coupled with the polymerase chain reaction (PCR) for the detection of a specific bacterial toxicity factor. 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 probe with a synthetic oligonucleotide was investigated; the absence of non-specific adsorption was verified using a non-complementary oligonucleotide. Many cycles of measurements can be performed on the same crystal surface by regenerating the single strand with 1 mM HCl. The same hybridisation reaction was then performed using real samples of DNA extracted from bacteria and amplified by PCR. The PCR product was a fragment of a specific gene of Aeromonas hydrophila. The piezoelectric biosensor was able to distinguish samples containing the gene or not; in this way it was possible to determine the pathogenicity of different Aeromonas strains isolated from water, vegetables or human specimens. Experiments with non-specific samples confirmed the absence of adsorption or non-specific effects on the quartz crystal treated with the reported procedure. (C) 2000 Elsevier Science B.V. All rights reserved.
Place, publisher, year, edition, pages
Elsevier Science B.V., Amsterdam. , 2000. Vol. 418, no 1
DNA piezoelectric biosensor; polymerase chain reaction; bacterial toxicity; hybridisation
Engineering and Technology
IdentifiersURN: urn:nbn:se:liu:diva-65236ISI: 000088275600001OAI: oai:DiVA.org:liu-65236DiVA: diva2:394998