Reliability of Pseudomonas aeruginosa semi-automated rep-PCR genotyping in various epidemiological situations

Doléans-Jordheim, Anne; Cournoyer, Benoît; Bergeron, Emmanuelle; Croizé, J.; Salord, Hélène; André, Jean; Mazoyer, Marie-Andrée; Renaud, François; Freney, J.

doi:10.1007/s10096-009-0755-z

Cited by 41 publications

(39 citation statements)

References 13 publications

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“…Regarding E. coli, DL presented good performance overall, except for occasional violations of MLST assignment. Our findings are in accordance with conclusions from a previous work that showed that DL is helpful for ruling out unrelated E. coli strains from (11,24). Finally, DL demonstrated excellent epidemiological concordance by correctly linking all outbreak-related strains of VREF, K. pneumoniae, A. baumannii, and P. aeruginosa in accordance with recent studies (12,16,24).…”

Section: Discussionsupporting

confidence: 92%

“…The DL system is a rapid and semiautomated rep-PCR commercial system that uses standard PCR kits and high-resolution amplicon detection by microfluidic capillary electrophoresis. Its discriminatory power and concordance with other typing methods have been previously evaluated for a number of bacterial pathogens (5,7,11,13,16,19,21,23,24). A recent study analyzed its usefulness in identifying hospital outbreaks caused by different bacterial pathogens (12).…”

Section: Discussionmentioning

confidence: 99%

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Controlled Performance Evaluation of the DiversiLab Repetitive-Sequence-Based Genotyping System for Typing Multidrug-Resistant Health Care-Associated Bacterial Pathogens

Deplano¹,

Denis²,

Rodríguez-Villalobos³

et al. 2011

J Clin Microbiol

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Fast, reliable, and versatile typing tools are essential to differentiate among related bacterial strains for epidemiological investigation and surveillance of health care-associated infection with multidrug-resistant (MDR) pathogens. The DiversiLab (DL) system is a semiautomated repetitive-sequence-based PCR system designed for rapid genotyping. The DL system performance was assessed by comparing its reproducibility, typeability, discriminatory power, and concordance with those of pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) and by assessing its epidemiological concordance on well-characterized MDR bacterial strains (n ‫؍‬ 165). These included vanA Enterococcus faecium, extended-spectrum ␤-lactamase (ESBL)-producing strains of Klebsiella pneumoniae, Escherichia coli, and Acinetobacter baumannii, and ESBL-or metallo-␤-lactamase (MBL)-producing Pseudomonas aeruginosa strains. The DL system showed very good performance for E. faecium and K. pneumoniae and good performance for other species, except for a discrimination index of <95% for A. baumannii and E. coli (93.9% and 93.5%, respectively) and incomplete concordance with MLST for P. aeruginosa (78.6%) and E. coli (97.0%). Occasional violations of MLST assignment by DL types were noted for E. coli. Complete epidemiological concordance was observed for all pathogens, as all outbreak-associated strains clustered in identical DL types that were distinct from those of unrelated strains. In conclusion, the DL system showed good to excellent performance, making it a reliable typing tool for investigation of outbreaks caused by study pathogens, even though it was generally less discriminating than PFGE analysis. For E. coli and P. aeruginosa, MLST cannot be reliably inferred from DL type due to phylogenetic group violation or discordance.Rapid assessment of microbial clonal relationships enables the tracking of the spread of multidrug-resistant (MDR) pathogens and guides transmission control measures in the hospital setting. The ideal typing method should be rapid, easy to perform, high throughput, and applicable to a wide range of microorganisms and should meet performance criteria, such as full typeability, reproducibility, high discriminatory power, and concordance with validated typing methods as well as consistency with underlying subspecies genetic population structures (29, 31). In the hospital setting, identification of epidemic strains is usually based on fingerprinting methods, such as pulsed-field gel electrophoresis (PFGE), a labor-intensive method that requires 3 to 4 days to produce results. The DiversiLab (DL) system (bioMérieux, Marcy l'Etoile, France) is a repetitive-sequence-based PCR (rep-PCR) technology which offers semiautomated, easy-to-use, high-throughput, and rapid bacterial strain typing. It could be a suitable alternative to PFGE analysis for outbreak investigation of health care-associated pathogens. This method was recently validated for typing of Acinetobacter spp., Escherichia coli, and Klebsiella spp., but...

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Section: Discussionsupporting

confidence: 92%

Section: Discussionmentioning

confidence: 99%

Controlled Performance Evaluation of the DiversiLab Repetitive-Sequence-Based Genotyping System for Typing Multidrug-Resistant Health Care-Associated Bacterial Pathogens

Deplano¹,

Denis²,

Rodríguez-Villalobos³

et al. 2011

J Clin Microbiol

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“…Several studies reported genetic diversity and heterogeneity among P. aeruginosa clinical isolates using ERIC-PCR, rep-PCR, PFGE, and RAPD-PCR methods in hospitals of Iran as well as other countries (7,16,17 aeruginosa isolates in hospitals especially burn centers.…”

Section: Discussionmentioning

confidence: 99%

Molecular Analysis of Pseudomonas aeruginosa Strains Isolated from Burn Patients by Repetitive Extragenic Palindromic-PCR (rep-PCR)

Sorkh

Shokoohizadeh

Rashidi

et al. 2017

Iran Red Crescent Med J

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Background: Infections caused by Pseudomonas aeruginosa raise an important issue in burn patients. Molecular epidemiologic studies have been used for investigating the genetic features of P. aeruginosa and rep-PCR technique has been introduced as a rapid low cost method. Objectives: This study focused on investigating the genetic similarity and antibiotic resistance pattern of P. aeruginosa isolated from the clinical samples of burn patients in a major burn center in Khuzestan Province, Iran. Methods: In a cross sectional study, a total of 75 strains of P. aeruginosa were isolated from burn patients at Taleghani hospital, which is the main burn center in Ahvaz, Iran, during May-September, 2015. Antimicrobial susceptibility of the isolates was detected using the disk diffusion method. Genetic relatedness of the isolates was analyzed by the rep-PCR technique. Results: Antimicrobial susceptibility testing showed more than 80% of P. aeruginosa isolates were resistant to ceftriaxone, cefotaxime, meropenem, piperacillin/tazobactam, ticarcillin, ciprofloxacin, and amikacin. Based on the rep-PCR analysis, 20 different common types and 20 unique patterns were illustrated among P. aeruginosa isolates. Conclusions: According to the findings of our study, there were diverse and high-level resistant P. aeruginosa strains in the major burn center in Khuzestan. Therefore, we faced troubles controlling the diverse P. aeruginosa clones in the burn patients.

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“…For isolates to be considered members of an MRPA clonal group, a threshold of 95% or greater similarity was used (17). The molecular profiles and dendrogram are displayed in Fig.…”

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confidence: 99%

Molecular Epidemiological Surveillance of Multidrug-Resistant Pseudomonas aeruginosa Isolates in a Pediatric Population of Patients with Cystic Fibrosis and Determination of Risk Factors for Infection with the Houston-1 Strain

et al. 2013

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e Multiyear molecular epidemiological surveillance of multidrug-resistant Pseudomonas aeruginosa (MRPA) in a pediatric cystic fibrosis care center identified an endemic MRPA strain (Houston-1). Recent hospitalization was found to be a statistically significant risk factor for acquisition of the endemic strain. Multiple infection control improvements led to the reduced incidence of the Houston-1 strain in the CF population.

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Reliability of Pseudomonas aeruginosa semi-automated rep-PCR genotyping in various epidemiological situations

Cited by 41 publications

References 13 publications

Controlled Performance Evaluation of the DiversiLab Repetitive-Sequence-Based Genotyping System for Typing Multidrug-Resistant Health Care-Associated Bacterial Pathogens

Controlled Performance Evaluation of the DiversiLab Repetitive-Sequence-Based Genotyping System for Typing Multidrug-Resistant Health Care-Associated Bacterial Pathogens

Molecular Analysis of Pseudomonas aeruginosa Strains Isolated from Burn Patients by Repetitive Extragenic Palindromic-PCR (rep-PCR)

Molecular Epidemiological Surveillance of Multidrug-Resistant Pseudomonas aeruginosa Isolates in a Pediatric Population of Patients with Cystic Fibrosis and Determination of Risk Factors for Infection with the Houston-1 Strain

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