Development of a Flow Cytometry-Based Method for Rapid Detection of Escherichia coli and Shigella Spp. Using an Oligonucleotide Probe

Autor: Fatemeh Rafii, Dan A. Buzatu, Yong Xue, Ted J. Moskal, Jon G. Wilkes, Rajesh Nayak, Anna J. Williams, Willie M. Cooper
Jazyk: angličtina
Rok vydání: 2016
Předmět:
0301 basic medicine
Bacterial Diseases
Luminescence
Light
Polymers
Oligonucleotides
lcsh:Medicine
medicine.disease_cause
Pathology and Laboratory Medicine
Foodborne Diseases
Scattering
Spectrum Analysis Techniques
Salmonella
RNA
Ribosomal
16S

Medicine and Health Sciences
Agar
Shigella
lcsh:Science
Escherichia coli Infections
Multidisciplinary
biology
Hybridization probe
Physics
Electromagnetic Radiation
Flow Cytometry
Bacterial Pathogens
Chemistry
Infectious Diseases
Salmonella Enterica
Macromolecules
Medical Microbiology
Spectrophotometry
Physical Sciences
Cytophotometry
Pathogens
Oligomer restriction
Research Article
food.ingredient
Materials by Structure
030106 microbiology
Cell Enumeration Techniques
Materials Science
Molecular Probe Techniques
Food Contamination
Escherichia coli O157
Research and Analysis Methods
Microbiology
Fluorescence
Agar plate
03 medical and health sciences
food
Enterobacteriaceae
medicine
Humans
Molecular Biology Techniques
Escherichia coli
Microbial Pathogens
Molecular Biology
Dysentery
Bacillary

Bacteria
lcsh:R
Organisms
Light Scattering
Biology and Life Sciences
biology.organism_classification
Polymer Chemistry
Molecular biology
Probe Hybridization
030104 developmental biology
lcsh:Q
Oligonucleotide Probes
Cytometry
Zdroj: PLoS ONE, Vol 11, Iss 2, p e0150038 (2016)
PLoS ONE
ISSN: 1932-6203
Popis: Standard methods to detect Escherichia coli contamination in food use the polymerase chain reaction (PCR) and agar culture plates. These methods require multiple incubation steps and take a long time to results. An improved rapid flow-cytometry based detection method was developed, using a fluorescence-labeled oligonucleotide probe specifically binding a16S rRNA sequence. The method positively detected 51 E. coli isolates as well as 4 Shigella species. All 27 non-E. coli strains tested gave negative results. Comparison of the new genetic assay with a total plate count (TPC) assay and agar plate counting indicated similar sensitivity, agreement between cytometry cell and colony counts. This method can detect a small number of E.coli cells in the presence of large numbers of other bacteria. This method can be used for rapid, economical, and stable detection of E. coli and Shigella contamination in the food industry and other contexts.
Databáze: OpenAIRE