A Comparison of Molecular Typing Methods Applied to Enterobacter cloacae complex: hsp60 Sequencing, Rep-PCR, and MLST

Molecular typing using repetitive sequenced-based PCR (rep-PCR) and hsp60 sequencing were applied to a collection of diverse Enterobacter cloacae complex isolates. To determine the most practical method for reference laboratories, we analyzed 71 E. cloacae complex isolates from sporadic and outbreak occurrences originating from 4 geographic areas. While rep-PCR was more discriminating, hsp60 sequencing provided a broader and a more objective geographical tracking method similar to multilocus sequence typing (MLST). In addition, we suggest that MLST may have higher discriminative power compared to hsp60 sequencing, although rep-PCR remains the most discriminative method for local outbreak investigations. In addition, rep-PCR can be an effective and inexpensive method for local outbreak investigation.

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Molecular typing of bacteria for epidemiological surveillance and outbreak investigation / Molekulare Typisierung von Bakterien für die epidemiologische Überwachung und Ausbruchsabklärung

Die Bodenkultur Journal of Land Management Food and Environment ◽

10.1515/boku-2016-0017 ◽

2016 ◽

Vol 67 (4) ◽

pp. 199-224 ◽

Cited By ~ 3

Author(s):

Werner Ruppitsch

Keyword(s):

Molecular Typing ◽

Pathogenic Bacteria ◽

Animal Health ◽

Epidemiological Surveillance ◽

Healthcare Facilities ◽

Microbial Infections ◽

Molekulare Typisierung ◽

Outbreak Investigations ◽

Typing Methods ◽

Phenotypic Methods

SummaryConstant confrontations with microbial threats pose major challenges to human and animal health, agricultural and food production, and public safety. Identifying pathogenic bacteria (species) and tracking strains (by series of well-characterized isolates) to their sources are especially important in outbreak investigations. Compared to the identification of the species, the identification of the source and spread of microbial infections represents a major—and many times futile—challenge. This is due to the multitude of ways microorganisms can occur and spread within healthcare facilities and in the community; how, when, and where they can contaminate the complex nutrition chain, leading to natural and man-made outbreaks.Typing is the characterization of isolates or strains below species or subspecies level. Typing of bacterial isolates is an essential procedure to identify the microbe causing the illness or to track down an outbreak to the suspected source. In the genomic era, the introduction of molecular methods has largely replaced phenotypic methods and “molecular epidemiology” has emerged as a new discipline. The current molecular typing methods can be classified into three categories: (a) PCR-based methods, (b) DNA fragment analysis-based methods, and (c) DNA sequence-based methods, including the new exciting era of high-throughput genome sequencing.

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Epidemiology and pathogenicity of Enterobacter cloacae complex clinical isolates recovered from bone and joint infections

10.26226/morressier.56d5ba30d462b80296c955c7 ◽

2016 ◽

Author(s):

Vincent Cattoir

Keyword(s):

Enterobacter Cloacae ◽

Clinical Isolates ◽

Joint Infections ◽

Bone And Joint Infections ◽

Enterobacter Cloacae Complex

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Outbreak ofStenotrophomonas maltophiliaBacteremia Among Patients Undergoing Bone Marrow Transplantation: Association With Faulty Replacement of Handwashing Soap

Infection Control and Hospital Epidemiology ◽

10.1086/501578 ◽

1999 ◽

Vol 20 (11) ◽

pp. 756-758 ◽

Cited By ~ 30

Author(s):

Jeffrey D. Klausner ◽

Carol Zukerman ◽

Ajit P. Limaye ◽

Lawrence Corey

Keyword(s):

Bone Marrow ◽

Bone Marrow Transplantation ◽

Healthcare Worker ◽

Molecular Typing ◽

Marrow Transplantation ◽

Stenotrophomonas Maltophilia ◽

Marrow Transplant ◽

Hand Washing ◽

Transplant Patients ◽

Typing Methods

AbstractUsing molecular typing methods, we confirmed an outbreak ofStenotrophomonas maltophiliaamong bone marrow transplant patients. The likely source was a healthcare worker who may have washed with moisturizer instead of soap between patients. Hospital epidemiologists need to go beyond antibiograms when evaluating outbreaks and be vigilant about all aspects of hand washing.

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Development of an ERIC sequence typing scheme for Laribacter hongkongensis, an emerging pathogen associated with community-acquired gastroenteritis and travellers’ diarrhoea

Journal of Medical Microbiology ◽

10.1099/jmm.0.049858-0 ◽

2013 ◽

Vol 62 (5) ◽

pp. 701-707 ◽

Cited By ~ 7

Author(s):

Jia-Li Feng ◽

Jin-Yan Lin ◽

Xiao-Bing Jiang ◽

Ou Zhang ◽

Jiang-Feng Zhu ◽

...

Keyword(s):

Clustering Analysis ◽

Consensus Sequence ◽

Pulsed Field Gel Electrophoresis ◽

Epidemiological Investigation ◽

Enterobacterial Repetitive Intergenic Consensus ◽

Emerging Pathogen ◽

Outbreak Investigations ◽

Laribacter Hongkongensis ◽

Specialized Equipment ◽

Typing Methods

Laribacter hongkongensis is a potential emerging pathogen, associated with community-acquired diarrhoea. For epidemiological purposes, different molecular typing methods, such as pulsed-field gel electrophoresis (PFGE) and multi-locus sequence typing, have been developed for this pathogen. However, these methods require specialized equipment and costly reagents. More importantly, they are labour-intensive and time-consuming, which is not really suitable for foodborne disease outbreak investigations. In this study, we developed a rapid and reliable method using 22-mer primers specific for the enterobacterial repetitive intergenic consensus sequence (ERIC). PFGE was used for comparison, to evaluate this method. A total of 81 isolates of L. hongkongensis were examined: 79 isolates recovered from food of diverse origins and two strains derived from patients with L. hongkongensis-associated infection. Typing patterns and clustering analysis indicated that the 81 L. hongkongensis isolates were grouped into 21 and 13 genotypes by ERIC-PCR and PFGE, respectively. ERIC-PCR was found as reproducible as PFGE. A high percentage (70.4 %) of isolates yielded distinguishable ERIC-PCR patterns, which were concordant with the results from PFGE. These results suggest that ERIC-PCR is valuable for use in the epidemiological investigation of L. hongkongensis.

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Draft Genome Sequences of Four Clinical Legionella pneumophila Isolates from Ontario, Canada

Genome Announcements ◽

10.1128/genomea.00295-18 ◽

2018 ◽

Vol 6 (15) ◽

pp. e00295-18

Author(s):

Alexander Fortuna ◽

Ricardo Ramnarine ◽

Aimin Li ◽

Nahuel Fittipaldi ◽

Christine Frantz ◽

...

Keyword(s):

Legionella Pneumophila ◽

Genetic Relationships ◽

Draft Genome ◽

Genome Sequences ◽

Content Type ◽

Outbreak Investigations ◽

Typing Methods

ABSTRACT Legionella pneumophila outbreak investigations require the development of reliable typing methods to better understand the genetic relationships of the isolates involved. Here, we report the draft genome sequences of four clinical Legionella pneumophila isolates obtained between 2000 and 2012 in Ontario, Canada.

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Putative Integrative Mobile Elements That Exploit the Xer Recombination Machinery Carrying blaIMI-Type Carbapenemase Genes in Enterobacter cloacae Complex Isolates in Singapore

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.01542-17 ◽

2017 ◽

Vol 62 (1) ◽

Cited By ~ 6

Author(s):

Tse H. Koh ◽

Nurdyana Binte Abdul Rahman ◽

Jeanette W. P. Teo ◽

My-Van La ◽

Balamurugan Periaswamy ◽

...

Keyword(s):

Phylogenetic Analysis ◽

Whole Genome Sequencing ◽

Genome Sequencing ◽

Multilocus Sequence Typing ◽

Enterobacter Cloacae ◽

Mobile Elements ◽

Whole Genome ◽

Content Type ◽

Flanking Regions ◽

Enterobacter Cloacae Complex

ABSTRACT Whole-genome sequencing was performed on 16 isolates of the carbapenemase-producing Enterobacter cloacae complex to determine the flanking regions of bla IMI-type genes. Phylogenetic analysis of multilocus sequence typing (MLST) targets separated the isolates into 4 clusters. The bla IMI-type genes were all found on Xer-dependent integrative mobile elements (IMEX). The IMEX elements of 5 isolates were similar to those described in Canada, while the remainder were novel. Five isolates had IMEX elements lacking a resolvase and recombinase.

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