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Towards Q-PCR of pathogenic bacteria with improved electrochemical double-tagged genosensing detection.
Biosens Bioelectron. 2008 Jul 15; 23(12):1805-11.BB

Abstract

A very sensitive assay for the rapid detection of pathogenic bacteria based on electrochemical genosensing has been designed. The assay was performed by the PCR specific amplification of the eaeA gene, related with the pathogenic activity of Escherichia coli O157:H7. The efficiency and selectivity of the selected primers were firstly studied by using standard Quantitative PCR (Q-PCR) based on TaqMan fluorescent strategy. The bacteria amplicon was detected by using two different electrochemical genosensing strategies, a highly selective biosensor based on a bulk-modified avidin biocomposite (Av-GEB) and a highly sensitive magneto sensor (m-GEC). The electrochemical detection was achieved in both cases by the enzyme marker HRP. The assay showed to be very sensitive, being able to detect 4.5 ng microl(-1) and 0.45 ng microl(-1) of the original bacterial genome after only 10 cycles of PCR amplification, when the first and the second strategies were used, respectively. Moreover, the electrochemical strategies for the detection of the amplicon showed to be more sensitive compared with Q-PCR strategies based on fluorescent labels such as TaqMan probes.

Authors+Show Affiliations

Departament de Química, Grup de Sensors i Biosensors, Universitat Autònoma de Barcelona, 08193 Bellaterra, Catalonia, Spain.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

18407486

Citation

Lermo, A, et al. "Towards Q-PCR of Pathogenic Bacteria With Improved Electrochemical Double-tagged Genosensing Detection." Biosensors & Bioelectronics, vol. 23, no. 12, 2008, pp. 1805-11.
Lermo A, Zacco E, Barak J, et al. Towards Q-PCR of pathogenic bacteria with improved electrochemical double-tagged genosensing detection. Biosens Bioelectron. 2008;23(12):1805-11.
Lermo, A., Zacco, E., Barak, J., Delwiche, M., Campoy, S., Barbé, J., Alegret, S., & Pividori, M. I. (2008). Towards Q-PCR of pathogenic bacteria with improved electrochemical double-tagged genosensing detection. Biosensors & Bioelectronics, 23(12), 1805-11. https://doi.org/10.1016/j.bios.2008.02.020
Lermo A, et al. Towards Q-PCR of Pathogenic Bacteria With Improved Electrochemical Double-tagged Genosensing Detection. Biosens Bioelectron. 2008 Jul 15;23(12):1805-11. PubMed PMID: 18407486.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Towards Q-PCR of pathogenic bacteria with improved electrochemical double-tagged genosensing detection. AU - Lermo,A, AU - Zacco,E, AU - Barak,J, AU - Delwiche,M, AU - Campoy,S, AU - Barbé,J, AU - Alegret,S, AU - Pividori,M I, Y1 - 2008/02/29/ PY - 2007/12/12/received PY - 2008/02/13/revised PY - 2008/02/21/accepted PY - 2008/4/15/pubmed PY - 2008/8/20/medline PY - 2008/4/15/entrez SP - 1805 EP - 11 JF - Biosensors & bioelectronics JO - Biosens Bioelectron VL - 23 IS - 12 N2 - A very sensitive assay for the rapid detection of pathogenic bacteria based on electrochemical genosensing has been designed. The assay was performed by the PCR specific amplification of the eaeA gene, related with the pathogenic activity of Escherichia coli O157:H7. The efficiency and selectivity of the selected primers were firstly studied by using standard Quantitative PCR (Q-PCR) based on TaqMan fluorescent strategy. The bacteria amplicon was detected by using two different electrochemical genosensing strategies, a highly selective biosensor based on a bulk-modified avidin biocomposite (Av-GEB) and a highly sensitive magneto sensor (m-GEC). The electrochemical detection was achieved in both cases by the enzyme marker HRP. The assay showed to be very sensitive, being able to detect 4.5 ng microl(-1) and 0.45 ng microl(-1) of the original bacterial genome after only 10 cycles of PCR amplification, when the first and the second strategies were used, respectively. Moreover, the electrochemical strategies for the detection of the amplicon showed to be more sensitive compared with Q-PCR strategies based on fluorescent labels such as TaqMan probes. SN - 0956-5663 UR - https://www.unboundmedicine.com/medline/citation/18407486/Towards_Q_PCR_of_pathogenic_bacteria_with_improved_electrochemical_double_tagged_genosensing_detection_ DB - PRIME DP - Unbound Medicine ER -