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Flow analysis coupled with PQC/DNA biosensor for assay of E. coli based on detecting DNA products from PCR amplification.
Biosens Bioelectron. 2006 Oct 15; 22(4):506-12.BB

Abstract

A flow-through PQC/DNA biosensor system is developed by combining sequential flow polymerase chain reaction (PCR) products denaturing prior to piezoelectric quartz crystal (PQC) detection via hybridization of ssDNA. The PQC/DNA biosensor is fabricated based on complex formation of neutravidin/biotinylated probe in 0.2M NaCl in TE buffer (10mM Tris, 1mM EDTA, pH 7.5). Results show that the coating fabricated provides a desirable quality with satisfactory performance. Its application for Escherichia coli detection under controlled flow at 0.02 mL/min for denaturing PCR products and 10 mL/min for transferring solution between reactors and delivering samples to detector to reduce rehybridization leads to significant improvement in repeatability (R.S.D.<6%, n=5) and sensitivity (DeltaF=34 Hz/1000 E. coli cells) as compared to existing manual method (R.S.D.=19%, n=5 and DeltaF=26 Hz/1000 E. coli cells, respectively). Down to 23 E. coli cells are detected, satisfying the HKEPD requirements for E. coli count in beach water.

Authors+Show Affiliations

Department of Chemistry, The University of Hong Kong, Pokfulam Road, Hong Kong SAR, China.No affiliation info availableNo affiliation info available

Pub Type(s)

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

Language

eng

PubMed ID

16971109

Citation

Sun, Hui, et al. "Flow Analysis Coupled With PQC/DNA Biosensor for Assay of E. Coli Based On Detecting DNA Products From PCR Amplification." Biosensors & Bioelectronics, vol. 22, no. 4, 2006, pp. 506-12.
Sun H, Zhang Y, Fung Y. Flow analysis coupled with PQC/DNA biosensor for assay of E. coli based on detecting DNA products from PCR amplification. Biosens Bioelectron. 2006;22(4):506-12.
Sun, H., Zhang, Y., & Fung, Y. (2006). Flow analysis coupled with PQC/DNA biosensor for assay of E. coli based on detecting DNA products from PCR amplification. Biosensors & Bioelectronics, 22(4), 506-12.
Sun H, Zhang Y, Fung Y. Flow Analysis Coupled With PQC/DNA Biosensor for Assay of E. Coli Based On Detecting DNA Products From PCR Amplification. Biosens Bioelectron. 2006 Oct 15;22(4):506-12. PubMed PMID: 16971109.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Flow analysis coupled with PQC/DNA biosensor for assay of E. coli based on detecting DNA products from PCR amplification. AU - Sun,Hui, AU - Zhang,Youyu, AU - Fung,Yingsing, Y1 - 2006/09/12/ PY - 2006/02/28/received PY - 2006/07/30/revised PY - 2006/08/09/accepted PY - 2006/9/15/pubmed PY - 2007/1/18/medline PY - 2006/9/15/entrez SP - 506 EP - 12 JF - Biosensors & bioelectronics JO - Biosens Bioelectron VL - 22 IS - 4 N2 - A flow-through PQC/DNA biosensor system is developed by combining sequential flow polymerase chain reaction (PCR) products denaturing prior to piezoelectric quartz crystal (PQC) detection via hybridization of ssDNA. The PQC/DNA biosensor is fabricated based on complex formation of neutravidin/biotinylated probe in 0.2M NaCl in TE buffer (10mM Tris, 1mM EDTA, pH 7.5). Results show that the coating fabricated provides a desirable quality with satisfactory performance. Its application for Escherichia coli detection under controlled flow at 0.02 mL/min for denaturing PCR products and 10 mL/min for transferring solution between reactors and delivering samples to detector to reduce rehybridization leads to significant improvement in repeatability (R.S.D.<6%, n=5) and sensitivity (DeltaF=34 Hz/1000 E. coli cells) as compared to existing manual method (R.S.D.=19%, n=5 and DeltaF=26 Hz/1000 E. coli cells, respectively). Down to 23 E. coli cells are detected, satisfying the HKEPD requirements for E. coli count in beach water. SN - 0956-5663 UR - https://www.unboundmedicine.com/medline/citation/16971109/Flow_analysis_coupled_with_PQC/DNA_biosensor_for_assay_of_E__coli_based_on_detecting_DNA_products_from_PCR_amplification_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0956-5663(06)00364-2 DB - PRIME DP - Unbound Medicine ER -