Details:

Alternative Title:
J Microbiol Methods
Personal Author:
Lindsey, Rebecca L. ; Garcia-Toledo, L. ; Fasulo, D. ; Gladney, L.M. ; Strockbine, N.
Lindsey, Rebecca L. ; Garcia-Toledo, L. ; Fasulo, D. ; Gladney, L.M. ; Strockbine, N. Less -
Description:
Escherichia coli, Escherichia albertii, and Escherichia fergusonii are closely related bacteria that can cause illness in humans, such as bacteremia, urinary tract infections and diarrhea. Current identification strategies for these three species vary in complexity and typically rely on the use of multiple phenotypic and genetic tests. To facilitate their rapid identification, we developed a multiplex PCR assay targeting conserved, species-specific genes. We used the Daydreamer™ (Pattern Genomics, USA) software platform to concurrently analyze whole genome sequence assemblies (WGS) from 150 Enterobacteriaceae genomes (107 E. coli, 5 Shigella spp., 21 E. albertii, 12 E. fergusonii and 5 other species) and design primers for the following species-specific regions: a 212bp region of the cyclic di-GMP regulator gene (cdgR, AW869_22935 from genome K-12 MG1655, CP014225) for E. coli/Shigella; a 393bp region of the DNA-binding transcriptional activator of cysteine biosynthesis gene (EAKF1_ch4033 from genome KF1, CP007025) for E. albertii; and a 575bp region of the palmitoleoyl-acyl carrier protein (ACP)-dependent acyltransferase (EFER_0790 from genome ATCC 35469, CU928158) for E. fergusonii. We incorporated the species-specific primers into a conventional multiplex PCR assay and assessed its performance with a collection of 97 Enterobacteriaceae strains. The assay was 100% sensitive and specific for detecting the expected species and offers a quick and accurate strategy for identifying E. coli, E. albertii, and E. fergusonii in either a single reaction or by in silico PCR with sequence assemblies.
Subjects:
[+]
Albertii Article Cross Infection DNA, Bacterial DNA Primers Enterobacteriaceae Escherichia Escherichia Coli Escherichia Coli Infections Escherichia Coli Proteins Fergusonii Genome, Bacterial Humans Multiplex Multiplex Polymerase Chain Reaction PCR Sensitivity And Specificity
Source:
J Microbiol Methods. 140:1-4
Pubmed ID:
28599915
Pubmed Central ID:
PMC5603207
Document Type:
Journal Article
Funding:
CC999999/Intramural CDC HHSUnited States/
Collection(s):
CDC Public Access
Main Document Checksum:
[+]
urn:sha256:cfab7da7d148dd01447aa8d20615c69718468396455ea828ca8ffcd3b192a4b2
Download URL:
https://stacks.cdc.gov/view/cdc/48230/cdc_48230_DS1.pdf
File Type:

Supporting Files

• 	NIHMS899984-supplement-supplementary_table_1.xlsx	xlsx
  	nihms899984.nxml	xml
  	nihms899984f1.gif	gif
  	nihms899984f1.jpg	jpeg
  	nihms899984f2.gif	gif
  	nihms899984f2.jpg	jpeg

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