Publication: Increased sensitivity of enterotoxigenic Escherichia coli detection in stool samples using oligonucleotide immobilized-magnetic nanoparticles
Issued Date
2021-12-01
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ISSN
2215017X
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2-s2.0-85116061368
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Mahidol University
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SCOPUS
Bibliographic Citation
Biotechnology Reports. Vol.32, (2021)
Suggested Citation
Kulachart Jangpatarapongsa, Kween Saimuang, Duangporn Polpanich, Raweewan Thiramanas, Mongkol Techakasikornpanich, Papichaya Yudech, Venusrin Paripurana, Chaniya Leepiyasakulchai, Pramuan Tangboriboonrat Increased sensitivity of enterotoxigenic Escherichia coli detection in stool samples using oligonucleotide immobilized-magnetic nanoparticles. Biotechnology Reports. Vol.32, (2021). doi:10.1016/j.btre.2021.e00677 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/75917
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Title
Increased sensitivity of enterotoxigenic Escherichia coli detection in stool samples using oligonucleotide immobilized-magnetic nanoparticles
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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.