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Increased sensitivity of enterotoxigenic Escherichia coli detection in stool samples using oligonucleotide immobilized-magnetic nanoparticles
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Document Title
Increased sensitivity of enterotoxigenic Escherichia coli detection in stool samples using oligonucleotide immobilized-magnetic nanoparticles
Author
Jangpatarapongsa K., Saimuang K., Polpanich D., Thiramanas R., Techakasikornpanich M., Yudech P., Paripurana V., Leepiyasakulchai C., Tangboriboonrat P.
Name from Authors Collection
Affiliations
Center for Research and Innovation, Faculty of Medical Technology, Mahidol University, Bangkok, 10700, Thailand; National Nanotechnology Center (NANOTEC), National Science and Technology Development Agency (NSTDA), Pathum Thani, 12120, Thailand; Department of Clinical Microbiology and Applied Technology, Faculty of Medical Technology, Mahidol UniversityBangkok 10700, Thailand; Department of Chemistry, Faculty of Science, Mahidol UniversityBangkok 10400, Thailand
Type
Article
Source Title
Biotechnology Reports
ISSN
2215017X
Year
2021
Volume
32
Open Access
All Open Access, Gold, Green
Publisher
Elsevier B.V.
DOI
10.1016/j.btre.2021.e00677
Format
Abstract
PCR detection of enterotoxigenic Escherichia-coli (ETEC) can be used directly on stool sample. However, it still has limitations due to presence of PCR inhibitors and interferences. This study, oligonucleotide primer specific to ETEC was immobilized onto MNPs and applied for separation and enrichment of ETEC-DNA from contaminants in stool after boiling. DNA separation efficiency was evaluated using conventional PCR and magneto-PCR-enzyme linked-gene-assay (MELGA). Due to high specificity of primer and efficiency of nanoparticles to bring down PCR inhibitors, DNA separation using primer-immobilized-MNPs exhibited 100-fold increase of sensitivity compared to that using simple boiling. Moreover, the sensitivities in stool were increased from 108 to 106 CFU/mL and 104 to 102 CFU/mL when PCR products were detected by gel electrophoresis and MELGA, respectively. Results suggested that oligonucleotide-immobilized-MNPs combined with boiling DNA extraction method was successfully used to separate the DNA of ETEC in stool with high sensitivity using MELGA. © 2021 The Authors
Industrial Classification
Knowledge Taxonomy Level 1
Knowledge Taxonomy Level 2
Knowledge Taxonomy Level 3
Funding Sponsor
Mahidol University; National Science and Technology Development Agency; Thailand Research Fund; Thailand Graduate Institute of Science and Technology
License
N/A
Rights
N/A
Publication Source
Scopus