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Alkaline phosphatase labeled SERS active sandwich immunoassay for detection of Escherichia coli.
Spectrochim Acta A Mol Biomol Spectrosc. 2018 Apr 05; 194:8-13.SA

Abstract

In this study, a sandwich immunoassay method utilizing enzymatic activity of alkaline phosphatase (ALP) on 5-bromo-4-chloro-3-indolyl phosphate (BCIP) for Escherichia coli (E. coli) detection was developed using surface enhanced Raman spectroscopy (SERS). For this purpose, spherical magnetic gold coated core-shell nanoparticles (MNPs-Au) and rod shape gold nanoparticles (Au-NRs) were synthesized and modified for immunomagnetic separation (IMS) of E. coli from the solution. In order to specify the developed method to ALP activity, Au-NRs were labeled with this enzyme. After successful construction of the immunoassay, BCIP substrate was added to produce the SERS-active product; 5-bromo-4-chloro-3-indole (BCI). A good linearity (R2=0.992) was established between the specific SERS intensity of BCI at 600cm-1 and logarithmic E. coli concentration in the range of 1.7×101-1.7×106cfumL-1. LOD and LOQ values were also calculated and found to be 10cfumL-1 and 30cfumL-1, respectively.

Authors+Show Affiliations

Department of Food Engineering, Faculty of Engineering, Hacettepe University, Beytepe, 06800 Ankara, Turkey.Department of Food Engineering, Faculty of Engineering, Hacettepe University, Beytepe, 06800 Ankara, Turkey.Department of Food Engineering, Faculty of Engineering, Hacettepe University, Beytepe, 06800 Ankara, Turkey.Department of Analytical Chemistry, Faculty of Pharmacy, Gazi University, 06330 Ankara, Turkey.Department of Biology, Science Faculty, Gazi University, 06500 Ankara, Turkey.Department of Food Engineering, Faculty of Engineering, Hacettepe University, Beytepe, 06800 Ankara, Turkey. Electronic address: ihb@hacettepe.edu.tr.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

29306060

Citation

Bozkurt, Akif Goktug, et al. "Alkaline Phosphatase Labeled SERS Active Sandwich Immunoassay for Detection of Escherichia Coli." Spectrochimica Acta. Part A, Molecular and Biomolecular Spectroscopy, vol. 194, 2018, pp. 8-13.
Bozkurt AG, Buyukgoz GG, Soforoglu M, et al. Alkaline phosphatase labeled SERS active sandwich immunoassay for detection of Escherichia coli. Spectrochim Acta A Mol Biomol Spectrosc. 2018;194:8-13.
Bozkurt, A. G., Buyukgoz, G. G., Soforoglu, M., Tamer, U., Suludere, Z., & Boyaci, I. H. (2018). Alkaline phosphatase labeled SERS active sandwich immunoassay for detection of Escherichia coli. Spectrochimica Acta. Part A, Molecular and Biomolecular Spectroscopy, 194, 8-13. https://doi.org/10.1016/j.saa.2017.12.057
Bozkurt AG, et al. Alkaline Phosphatase Labeled SERS Active Sandwich Immunoassay for Detection of Escherichia Coli. Spectrochim Acta A Mol Biomol Spectrosc. 2018 Apr 5;194:8-13. PubMed PMID: 29306060.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Alkaline phosphatase labeled SERS active sandwich immunoassay for detection of Escherichia coli. AU - Bozkurt,Akif Goktug, AU - Buyukgoz,Guluzar Gorkem, AU - Soforoglu,Mehmet, AU - Tamer,Ugur, AU - Suludere,Zekiye, AU - Boyaci,Ismail Hakki, Y1 - 2017/12/21/ PY - 2017/09/14/received PY - 2017/12/09/revised PY - 2017/12/20/accepted PY - 2018/1/7/pubmed PY - 2018/8/18/medline PY - 2018/1/7/entrez KW - Alkaline phosphatase KW - Escherichia coli detection KW - Sandwich immunoassay KW - Surface enhanced Raman spectroscopy SP - 8 EP - 13 JF - Spectrochimica acta. Part A, Molecular and biomolecular spectroscopy JO - Spectrochim Acta A Mol Biomol Spectrosc VL - 194 N2 - In this study, a sandwich immunoassay method utilizing enzymatic activity of alkaline phosphatase (ALP) on 5-bromo-4-chloro-3-indolyl phosphate (BCIP) for Escherichia coli (E. coli) detection was developed using surface enhanced Raman spectroscopy (SERS). For this purpose, spherical magnetic gold coated core-shell nanoparticles (MNPs-Au) and rod shape gold nanoparticles (Au-NRs) were synthesized and modified for immunomagnetic separation (IMS) of E. coli from the solution. In order to specify the developed method to ALP activity, Au-NRs were labeled with this enzyme. After successful construction of the immunoassay, BCIP substrate was added to produce the SERS-active product; 5-bromo-4-chloro-3-indole (BCI). A good linearity (R2=0.992) was established between the specific SERS intensity of BCI at 600cm-1 and logarithmic E. coli concentration in the range of 1.7×101-1.7×106cfumL-1. LOD and LOQ values were also calculated and found to be 10cfumL-1 and 30cfumL-1, respectively. SN - 1873-3557 UR - https://www.unboundmedicine.com/medline/citation/29306060/Alkaline_phosphatase_labeled_SERS_active_sandwich_immunoassay_for_detection_of_Escherichia_coli_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1386-1425(17)31030-2 DB - PRIME DP - Unbound Medicine ER -