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| Titel |
New quantitative detection of pathogens in heterogeneous environmental samples |
| VerfasserIn |
Eun-Hee Lee, Xiaofang Wang, Kristi Mitchell, Seon-Ha Chae, Ahjeong Son |
| Konferenz |
EGU General Assembly 2015
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| Medientyp |
Artikel
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| Sprache |
Englisch
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| Digitales Dokument |
PDF |
| Erschienen |
In: GRA - Volume 17 (2015) |
| Datensatznummer |
250108215
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| Publikation (Nr.) |
 EGU/EGU2015-7957.pdf |
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| Zusammenfassung |
| Quantum dots and magnetic beads based genomic assay (NanoGene assay) has been
developed for sensitive and inhibition resistant gene quantification to achieve in-situ bacteria
monitoring in environmental samples. In this study, eaeA gene of pathogenic E. coli
O157:H7 was quantified. The result demonstrated the excellent sensitivity (i.e.,
limit of detection: 87 gene copies for dsDNA and 890 zeptomolar for ssDNA) in
the presence of nonspecific microbial populations (Kim et al., 2010; 2011a). The
feasibility of the developed gene quantification for non-laboratory environment usage
(in-situ use) was investigated. Therefore, DNA hybridization was achieved at ambient
temperature and minimum agitation, and the analysis was completed within hours. Most
importantly, the NanoGene assay demonstrated the resistance to the presence of
naturally occurring inhibitors (humic acids, cations) and residual reagents (surfactants,
alcohols) from DNA extraction (Kim et al., 2011b). The assay was also applied to
humic acids laden soils (7 types of soils with various amount of organic matters) and
successfully quantified 105 to 108 CFU of E. coli O157:H7 per gram soil (R2 =
0.99). The results indicate that the presented NanoGene assay is suitable for further
development as an in-situ bacteria monitoring method for working with heterogeneous
environmental samples (Wang et al., 2013). Another aspect of the method is to
transform the NanoGene assay into a portable device that can be used as a pathogenic
bacteria detector in environment. The project consisted of the first inline fluidic
components development and characterization as well as the first integration effort on a
briefcase platform for the in-situ pathogen detection system (IPDS) (Mitchell et
al., 2014). Our long term vision is to further miniaturize the briefcase platform
implementation of the IPDS and to commercialize the handheld version of the IPDS. |
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