Open AccessDevelopment of a Flow Cytometry-Based Method for Rapid Detection of Escherichia coli and Shigella Spp. Using an Oligonucleotide Probe
Author(s) -
Yong Xue,
Jon G. Wilkes,
Ted J. Moskal,
A. J. Williams,
Willie M. Cooper,
Rajesh Nayak,
Fatemeh Rafii,
Dan A. Buzatu
Publication year - 2016
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0150038
Subject(s) - shigella , escherichia coli , flow cytometry , biology , oligomer restriction , polymerase chain reaction , microbiology and biotechnology , agar , oligonucleotide , enterobacteriaceae , bacteria , enumeration , agar plate , dna , genetics , gene , mathematics , combinatorics
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.