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Source tracking of Escherichia coli by 16S-23S intergenic spacer region denaturing gradient gel electrophoresis (DGGE) of the rrnB ribosomal operon.
Can J Microbiol. 2007 Oct; 53(10):1174-84.CJ

Abstract

This research validates a novel approach for source tracking based on denaturing gradient gel electrophoresis (DGGE) analysis of DNA extracted from Escherichia coli isolates. Escherichia coli from different animal sources and from river samples upstream from, at, and downstream of a combined sewer overflow were subjected to DGGE to determine sequence variations within the 16S-23S intergenic spacer region (ISR) of the rrnB ribosomal operon. The ISR was analyzed to determine if E. coli isolates from various animal sources could be differentiated from each other. DNA isolated from the E. coli animal sources was PCR amplified to isolate the rrnB operon. To prevent amplification of all 7 E. coli ribosomal operons by PCR amplification using universal primers, sequence-specific primers were utilized for the rrnB operon. Another primer set was then used to prepare samples of the 16S-23S ISR for DGGE. Comparison of PCR-DGGE results between human and animal sources revealed differences in the distribution and frequency of the DGGE bands produced. Human and Canada Goose isolates had the most unique distribution patterns and the highest percent of unique isolates and were grouped separately from all other animal sources. Method validation suggests that there are enough host specificity and genetic differences for use in the field. Field results at and around a combined sewer overflow indicate that this method can be used for microbial source tracking.

Authors+Show Affiliations

Department of Biological Sciences, Youngstown State University, Youngstown, OH 44555, USA.No affiliation info availableNo affiliation info available

Pub Type(s)

Evaluation Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

18026210

Citation

D'Elia, Thomas V., et al. "Source Tracking of Escherichia Coli By 16S-23S Intergenic Spacer Region Denaturing Gradient Gel Electrophoresis (DGGE) of the rrnB Ribosomal Operon." Canadian Journal of Microbiology, vol. 53, no. 10, 2007, pp. 1174-84.
D'Elia TV, Cooper CR, Johnston CG. Source tracking of Escherichia coli by 16S-23S intergenic spacer region denaturing gradient gel electrophoresis (DGGE) of the rrnB ribosomal operon. Can J Microbiol. 2007;53(10):1174-84.
D'Elia, T. V., Cooper, C. R., & Johnston, C. G. (2007). Source tracking of Escherichia coli by 16S-23S intergenic spacer region denaturing gradient gel electrophoresis (DGGE) of the rrnB ribosomal operon. Canadian Journal of Microbiology, 53(10), 1174-84.
D'Elia TV, Cooper CR, Johnston CG. Source Tracking of Escherichia Coli By 16S-23S Intergenic Spacer Region Denaturing Gradient Gel Electrophoresis (DGGE) of the rrnB Ribosomal Operon. Can J Microbiol. 2007;53(10):1174-84. PubMed PMID: 18026210.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Source tracking of Escherichia coli by 16S-23S intergenic spacer region denaturing gradient gel electrophoresis (DGGE) of the rrnB ribosomal operon. AU - D'Elia,Thomas V, AU - Cooper,Chester R, AU - Johnston,Carl G, PY - 2007/11/21/pubmed PY - 2008/1/3/medline PY - 2007/11/21/entrez SP - 1174 EP - 84 JF - Canadian journal of microbiology JO - Can J Microbiol VL - 53 IS - 10 N2 - This research validates a novel approach for source tracking based on denaturing gradient gel electrophoresis (DGGE) analysis of DNA extracted from Escherichia coli isolates. Escherichia coli from different animal sources and from river samples upstream from, at, and downstream of a combined sewer overflow were subjected to DGGE to determine sequence variations within the 16S-23S intergenic spacer region (ISR) of the rrnB ribosomal operon. The ISR was analyzed to determine if E. coli isolates from various animal sources could be differentiated from each other. DNA isolated from the E. coli animal sources was PCR amplified to isolate the rrnB operon. To prevent amplification of all 7 E. coli ribosomal operons by PCR amplification using universal primers, sequence-specific primers were utilized for the rrnB operon. Another primer set was then used to prepare samples of the 16S-23S ISR for DGGE. Comparison of PCR-DGGE results between human and animal sources revealed differences in the distribution and frequency of the DGGE bands produced. Human and Canada Goose isolates had the most unique distribution patterns and the highest percent of unique isolates and were grouped separately from all other animal sources. Method validation suggests that there are enough host specificity and genetic differences for use in the field. Field results at and around a combined sewer overflow indicate that this method can be used for microbial source tracking. SN - 0008-4166 UR - https://www.unboundmedicine.com/medline/citation/18026210/Source_tracking_of_Escherichia_coli_by_16S_23S_intergenic_spacer_region_denaturing_gradient_gel_electrophoresis__DGGE__of_the_rrnB_ribosomal_operon_ L2 - https://cdnsciencepub.com/doi/10.1139/W07-083?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub=pubmed DB - PRIME DP - Unbound Medicine ER -