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Increased sensitivity of enterotoxigenic Escherichia coli detection in stool samples using oligonucleotide immobilized-magnetic nanoparticles

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 enrichmen...

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Autores principales: Jangpatarapongsa, Kulachart, Saimuang, Kween, Polpanich, Duangporn, Thiramanas, Raweewan, Techakasikornpanich, Mongkol, Yudech, Papichaya, Paripurana, Venusrin, Leepiyasakulchai, Chaniya, Tangboriboonrat, Pramuan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8487978/
https://www.ncbi.nlm.nih.gov/pubmed/34631437
http://dx.doi.org/10.1016/j.btre.2021.e00677
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author Jangpatarapongsa, Kulachart
Saimuang, Kween
Polpanich, Duangporn
Thiramanas, Raweewan
Techakasikornpanich, Mongkol
Yudech, Papichaya
Paripurana, Venusrin
Leepiyasakulchai, Chaniya
Tangboriboonrat, Pramuan
author_facet Jangpatarapongsa, Kulachart
Saimuang, Kween
Polpanich, Duangporn
Thiramanas, Raweewan
Techakasikornpanich, Mongkol
Yudech, Papichaya
Paripurana, Venusrin
Leepiyasakulchai, Chaniya
Tangboriboonrat, Pramuan
author_sort Jangpatarapongsa, Kulachart
collection PubMed
description 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 10(8) to 10(6) CFU/mL and 10(4) to 10(2) 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.
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spelling pubmed-84879782021-10-08 Increased sensitivity of enterotoxigenic Escherichia coli detection in stool samples using oligonucleotide immobilized-magnetic nanoparticles Jangpatarapongsa, Kulachart Saimuang, Kween Polpanich, Duangporn Thiramanas, Raweewan Techakasikornpanich, Mongkol Yudech, Papichaya Paripurana, Venusrin Leepiyasakulchai, Chaniya Tangboriboonrat, Pramuan Biotechnol Rep (Amst) Research Article 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 10(8) to 10(6) CFU/mL and 10(4) to 10(2) 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. Elsevier 2021-09-24 /pmc/articles/PMC8487978/ /pubmed/34631437 http://dx.doi.org/10.1016/j.btre.2021.e00677 Text en © 2021 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Research Article
Jangpatarapongsa, Kulachart
Saimuang, Kween
Polpanich, Duangporn
Thiramanas, Raweewan
Techakasikornpanich, Mongkol
Yudech, Papichaya
Paripurana, Venusrin
Leepiyasakulchai, Chaniya
Tangboriboonrat, Pramuan
Increased sensitivity of enterotoxigenic Escherichia coli detection in stool samples using oligonucleotide immobilized-magnetic nanoparticles
title Increased sensitivity of enterotoxigenic Escherichia coli detection in stool samples using oligonucleotide immobilized-magnetic nanoparticles
title_full Increased sensitivity of enterotoxigenic Escherichia coli detection in stool samples using oligonucleotide immobilized-magnetic nanoparticles
title_fullStr Increased sensitivity of enterotoxigenic Escherichia coli detection in stool samples using oligonucleotide immobilized-magnetic nanoparticles
title_full_unstemmed Increased sensitivity of enterotoxigenic Escherichia coli detection in stool samples using oligonucleotide immobilized-magnetic nanoparticles
title_short Increased sensitivity of enterotoxigenic Escherichia coli detection in stool samples using oligonucleotide immobilized-magnetic nanoparticles
title_sort increased sensitivity of enterotoxigenic escherichia coli detection in stool samples using oligonucleotide immobilized-magnetic nanoparticles
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8487978/
https://www.ncbi.nlm.nih.gov/pubmed/34631437
http://dx.doi.org/10.1016/j.btre.2021.e00677
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