Buckyballs conjugated with nucleic acid sequences identifies microorganisms in live cell assays
| Autor: | Bahram Parvin, Qingsu Cheng |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
lcsh:Medical technology Microorganism Point-of-Care Systems lcsh:Biotechnology 030106 microbiology Biomedical Engineering Pharmaceutical Science Medicine (miscellaneous) Bioengineering Bacillus subtilis Cell Separation Applied Microbiology and Biotechnology Sensitivity and Specificity 03 medical and health sciences RNA Ribosomal 16S lcsh:TP248.13-248.65 Fluorescence microscope 16S rRNA Microbial live cell assays Base Pairing Fluorescent Dyes biology Chemotactic Factors Research fungi Nucleic acid sequence food and beverages Aptamers Nucleotide Microfluidic Analytical Techniques biology.organism_classification Molecular biology Yeast C60 Streptococcus sanguinis 030104 developmental biology Biochemistry Microscopy Fluorescence lcsh:R855-855.5 Pseudomonas aeruginosa Nucleic acid Molecular Medicine Biological Assay Fullerenes Streptococcus sanguis Bacteria |
| Zdroj: | Journal of Nanobiotechnology, Vol 15, Iss 1, Pp 1-11 (2017) Journal of Nanobiotechnology |
| ISSN: | 1477-3155 |
| DOI: | 10.1186/s12951-017-0315-0 |
| Popis: | Background Rapid identification of bacteria can play an important role at the point of care, evaluating the health of the ecosystem, and discovering spatiotemporal distributions of a bacterial community. We introduce a method for rapid identification of bacteria in live cell assays based on cargo delivery of a nucleic acid sequence and demonstrate how a mixed culture can be differentiated using a simple microfluidic system. Methods C60 Buckyballs are functionalized with nucleic acid sequences and a fluorescent reporter to show that a diversity of microorganisms can be detected and identified in live cell assays. The nucleic acid complexes include an RNA detector, targeting a species-specific sequence in the 16S rRNA, and a complementary DNA with an attached fluorescent reporter. As a result, each bacterium can be detected and visualized at a specific emission frequency through fluorescence microscopy. Results The C60 probe complexes can detect and identify a diversity of microorganisms that include gram-position and negative bacteria, yeast, and fungi. More specifically, nucleic-acid probes are designed to identify mixed cultures of Bacillus subtilis and Streptococcus sanguinis, or Bacillus subtilis and Pseudomonas aeruginosa. The efficiency, cross talk, and accuracy for the C60 probe complexes are reported. Finally, to demonstrate that mixed cultures can be separated, a microfluidic system is designed that connects a single source-well to multiple sinks wells, where chemo-attractants are placed in the sink wells. The microfluidic system allows for differentiating a mixed culture. Conclusions The technology allows profiling of bacteria composition, at a very low cost, for field studies and point of care. Electronic supplementary material The online version of this article (10.1186/s12951-017-0315-0) contains supplementary material, which is available to authorized users. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
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