scholarly journals Effective Surveillance Using Multilocus Variable-Number Tandem-Repeat Analysis and Whole-Genome Sequencing for Enterohemorrhagic Escherichia coli O157

2019 ◽  
Vol 85 (17) ◽  
Author(s):  
Kenichi Lee ◽  
Hidemasa Izumiya ◽  
Sunao Iyoda ◽  
Makoto Ohnishi
Keyword(s):  
Escherichia Coli ◽  
Tandem Repeat ◽  
Core Genome ◽  
Variable Number Tandem Repeat ◽  
Variable Number ◽  
Whole Genome ◽  
Single Nucleotide ◽  
Molecular Surveillance ◽  
Content Type ◽  
Repeat Analysis

ABSTRACT Due to the potential of enterohemorrhagic Escherichia coli (EHEC) serogroup O157 to cause large food borne outbreaks, national and international surveillance is necessary. For developing an effective method of molecular surveillance, a conventional method, multilocus variable-number tandem-repeat analysis (MLVA), and whole-genome sequencing (WGS) analysis were compared. WGS of 369 isolates of EHEC O157 belonging to 7 major MLVA types and their relatives were subjected to comprehensive in silico typing, core genome single nucleotide polymorphism (cgSNP), and core genome multilocus sequence typing (cgMLST) analyses. The typing resolution was the highest in cgSNP analysis. However, determination of the sequence of the mismatch repair protein gene mutS is necessary because spontaneous deletion of the gene could lead to a hypermutator phenotype. MLVA had sufficient typing resolution for a short-term outbreak investigation and had advantages in rapidity and high throughput. cgMLST showed less typing resolution than cgSNP, but it is less time-consuming and does not require as much computer power. Therefore, cgMLST is suitable for comparisons using large data sets (e.g., international comparison using public databases). In conclusion, screening using MLVA followed by cgMLST and cgSNP analyses would provide the highest typing resolution and improve the accuracy and cost-effectiveness of EHEC O157 surveillance. IMPORTANCE Intensive surveillance for enterohemorrhagic Escherichia coli (EHEC) serogroup O157 is important to detect outbreaks and to prevent the spread of the bacterium. Recent advances in sequencing technology made molecular surveillance using whole-genome sequence (WGS) realistic. To develop rapid, high-throughput, and cost-effective typing methods for real-time surveillance, typing resolution of WGS and a conventional typing method, multilocus variable-number tandem-repeat analysis (MLVA), was evaluated. Nation-level systematic comparison of MLVA, core genome single nucleotide polymorphism (cgSNP), and core genome multilocus sequence typing (cgMLST) indicated that a combination of WGS and MLVA is a realistic approach to improve EHEC O157 surveillance.

scholarly journals Multiple-locus variable-number tandem repeat analysis for strain discrimination of non-O157 Shiga toxin-producing Escherichia coli

2016 ◽  
Vol 125 ◽  
pp. 70-80 ◽  
Author(s):  
Chris Timmons ◽  
Eija Trees ◽  
Efrain M. Ribot ◽  
Peter Gerner-Smidt ◽  
Patti LaFon ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Tandem Repeat ◽  
Shiga Toxin ◽  
Variable Number Tandem Repeat ◽  
Variable Number ◽  
Multiple Locus ◽  
Repeat Analysis

Microbiological Effects of a Routine Treatment for Decontaminating Hide-On Carcasses at a Large Beef Packing Plant

2015 ◽  
Vol 78 (2) ◽  
pp. 256-263 ◽  
Author(s):  
XIANQIN YANG ◽  
MADHU BADONI ◽  
FRANCES TRAN ◽  
COLIN O. GILL
Keyword(s):  
Escherichia Coli ◽  
Tandem Repeat ◽  
Variable Number Tandem Repeat ◽  
Variable Number ◽  
Routine Treatment ◽  
Multiple Locus ◽  
E Coli ◽  
Repeat Analysis ◽  
Before And After

To investigate the microbiological effects of a hide-on carcass decontaminating treatment recently implemented at a beef packing plant, carcasses undergoing routine processing at the plant were sampled during successive periods in January/February, April/May, and September/October. During each period, samples were collected from carcasses before and after the decontamination of hide-on carcasses, after skinning, before decontamination of the skinned carcasses, and at the end of the carcass dressing process. At each stage of processing during each period, samples were obtained by swabbing an area of 1,000 cm2 on each of 25 carcasses. Aerobes, coliforms, and Escherichia coli were enumerated. In most samples, coliforms were predominantly E. coli. In all three periods, the log mean numbers of aerobes and E. coli recovered from hides before decontamination were between 6.6 and 6.8 and between 5.3 and 5.9 log CFU/1,000 cm2, respectively. The log mean numbers of aerobes recovered from decontaminated hides were 6.6 log CFU/1,000 cm2 in January/February and April/May but 5.4 log CFU/1,000 cm2 in September/October. The log total numbers of E. coli recovered from decontaminated hides in January/February and April/May were 2.4 and 3.8 log CFU/25,000 cm2, respectively, but no E. coli was recovered from such carcasses in September/October. Log total numbers of aerobes and E. coli recovered from skinned or dressed carcasses were mostly >4 and between 1 and 2 log CFU/25,000 cm2, respectively. Typing of 480 E. coli isolates by multiple-locus variable-number tandem repeat analysis (MLVA) identified 218 MLVA types. Most isolates recovered from carcasses in different periods or at different stages of processing were of different MLVA types. However, small numbers of MLVA types were recovered in more than one period or from both hides before and after decontamination and skinned or dressed carcasses. The findings show that the hide-decontaminating treatment disrupted the usual transfer of E. coli from hides to meat surfaces during carcass skinning.

Inter-laboratory comparison of multi-locus variable-number tandem repeat analysis (MLVA) for verocytotoxin-producing Escherichia coli O157 to facilitate data sharing

2014 ◽  
Vol 143 (1) ◽  
pp. 104-107 ◽  
Author(s):  
A. HOLMES ◽  
N. PERRY ◽  
G. WILLSHAW ◽  
M. HANSON ◽  
L. ALLISON
Keyword(s):  
Escherichia Coli ◽  
Tandem Repeat ◽  
Variable Number Tandem Repeat ◽  
Variable Number ◽  
Escherichia Coli O157 ◽  
Consensus Guidelines ◽  
International Consensus ◽  
Repeat Analysis ◽  
Repeat Units ◽  
International Consensus Guidelines

SUMMARYMulti-locus variable number tandem repeat analysis (MLVA) is used in clinical and reference laboratories for subtyping verocytotoxin-producing Escherichia coli O157 (VTEC O157). However, as yet there is no common allelic or profile nomenclature to enable laboratories to easily compare data. In this study, we carried out an inter-laboratory comparison of an eight-loci MLVA scheme using a set of 67 isolates of VTEC O157. We found all but two isolates were identical in profile in the two laboratories, and repeat units were homogeneous in size but some were incomplete. A subset of the isolates (n = 17) were sequenced to determine the actual copy number of representative alleles, thereby enabling alleles to be named according to international consensus guidelines. This work has enabled us to realize the potential of MLVA as a portable, highly discriminatory and convenient subtyping method.

scholarly journals Multilocus Variable-Number Tandem Repeat Analysis Distinguishes Outbreak and Sporadic Escherichia coli O157:H7 Isolates

2004 ◽  
Vol 42 (2) ◽  
pp. 940-940
Author(s):  
A. C. Noller ◽  
M. C. McEllistrem ◽  
A. G. F. Pacheco ◽  
D. J. Boxrud ◽  
L. H. Harrison
Keyword(s):  
Escherichia Coli ◽  
Tandem Repeat ◽  
Variable Number Tandem Repeat ◽  
Variable Number ◽  
Escherichia Coli O157 ◽  
Repeat Analysis
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