| Single-cell detection: test of microbial contamination using capillary electrophoresis. | |
| | |
MedLine Citation:
|
PMID: 17297979 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
|
Single cells of bacteria and fungi were detected using a capillary electrophoresis-based test for microbial contamination in laboratory samples. This technique utilizes a dilute cationic surfactant buffer to sweep microorganisms out of the sample zone and a small plug of "blocking agent" to negate the cells' mobility and induce aggregation. Analysis times are generally under 10 min. Previously, a nutrient broth media was reported as an effective blocking agent; however, the natural background fluorescence from the nutrient broth limited the detection sensitivity to approximately 50 cells. In order to enhance the sensitivity of the technique down to a single cell, an alternative synthetic blocking agent was sought. Various potential blocking agents were screened including salts, polypeptides, small organic zwitterions, and surfactants. Zwitterionic surfactants are shown to be attractive alternatives to a nutrient broth blocker and mimic the nutrient broth's effects on cellular aggregation and mobility. Specifically, caprylyl sulfobetaine provided the sharpest cell peaks. By substituting caprylyl sulfobetaine in place of the nutrient broth, the fluorescence of the blocker plug was reduced by as much as 40 x. This reduction in background noise enables detection of a single microorganism in a sample and allows this technique to be potentially used as a rapid sterility test. All single cells analyzed using this technique displayed signal-to-noise ratios between 5 and 9. |
| | |
Authors:
|
Andrew W Lantz; Ye Bao; Daniel W Armstrong |
Related Documents
:
|
11226479 - Quantitative analysis of specific mrna species in minute cell samples by rt-pcr and flo... 9589579 - Flow cytometry data included in first round cluster analysis. 2458829 - Flow cytometric localization within the cell cycle and isolation of viable cells follow... 12776929 - A fluorescence microplate assay using yopro-1 to measure apoptosis: application to hl60... 15921499 - Isolation of the choanocyte in the fresh water sponge, ephydatia fluviatilis and its li... 15288249 - Flow cytometry for high-throughput, high-content screening. 3930709 - Photoperiodic variations of somatic and germ cell populations in the soay ram testis. 5864849 - Dielectric behavior of yeast cells treated with hgc-12 and cetyl trimethyl ammonium bro... 3969079 - Effects of x-irradiation on cell-cycle progression, induction of chromosomal aberration... |
Publication Detail:
|
Type: Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't |
Journal Detail:
|
Title: Analytical chemistry Volume: 79 ISSN: 0003-2700 ISO Abbreviation: Anal. Chem. Publication Date: 2007 Feb |
Date Detail:
|
Created Date: 2007-02-14 Completed Date: 2007-04-12 Revised Date: - |
Medline Journal Info:
|
Nlm Unique ID: 0370536 Medline TA: Anal Chem Country: United States |
Other Details:
|
Languages: eng Pagination: 1720-4 Citation Subset: IM |
Affiliation:
|
Iowa State University, Chemistry Department, 1605 Gilman Hall, Ames, Iowa 50011, USA. |
Export Citation:
|
APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
|
Bacteria
/
chemistry* Electrophoresis, Capillary Equipment Contamination Fungi / chemistry* Sensitivity and Specificity Surface-Active Agents / chemistry Time Factors |
| Chemical | |
Reg. No./Substance:
|
0/Surface-Active Agents |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Previous Document: Gel electrophoresis coupled to inductively coupled plasma-mass spectrometry using species-specific i...
Next Document: Microfluidic device for the detection of glucose using a micro direct methanol fuel cell as an amper...