A new highly discriminatory multiplex capillary-based MLVA assay as a tool for the epidemiological survey of Pseudomonas aeruginosa in cystic fibrosis patients

Sobral, Daniel; Mariani‐Kurkdjian, Patricia; Bingen, Édouard; Vu-Thien, Hoang; Hormigos, Katia; Lebeau, B.; Loisy-Hamon, Fabienne; Munck, À.; Vergnaud, Gilles; Pourcel, Christine

doi:10.1007/s10096-012-1562-5

Cited by 32 publications

(27 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Besides, the MLVA-10 scheme could easily be automated using fluorescent primers and a single multiplex PCR (31). Capillary electrophoresis would allow automatic size measurement of the PCR products and thus increase the accuracy of repeat number determination, as described for other bacterial species (31)(32)(33). Furthermore, some potentially informative VNTR markers that were ruled out from the present scheme due to short repeats (Ͻ15 bp) could be included to constitute a second panel with high discriminatory power, as has been proposed for other bacteria (34).…”

Section: Discussionmentioning

confidence: 99%

Development of a Multiple-Locus Variable-Number Tandem-Repeat Typing Scheme for Genetic Fingerprinting of Burkholderia cenocepacia and Application to Nationwide Epidemiological Analysis

Segonds¹,

Thouverez²,

Barthe³

et al. 2015

J Clin Microbiol

View full text Add to dashboard Cite

Organisms of the Burkholderia cepacia complex are especially important pathogens in cystic fibrosis (CF), with a propensity for patient-to-patient spread and long-term respiratory colonization. B. cenocepacia and Burkholderia multivorans account for the majority of infections in CF, and major epidemic clones have been recognized throughout the world. The aim of the present study was to develop and evaluate a multilocus variable-number tandem-repeat (VNTR) analysis (MLVA) scheme for B. cenocepacia. Potential VNTR loci were identified upon analysis of the annotated genome sequences of B. cenocepacia strains AU1054, J2315, and MCO-3, and 10 of them were selected on the basis of polymorphisms and size. A collection of 100 B. cenocepacia strains, including epidemiologically related and unrelated strains, as well as representatives of the major epidemic lineages, was used to evaluate typeability, epidemiological concordance, and the discriminatory power of MLVA-10 compared with those of pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). Longitudinal stability was assessed by testing 39 successive isolates from 14 patients. Typeability ranged from 0.91 to 1, except for that of one marker, which was not amplified in 53% of the B. cenocepacia IIIA strains. The MLVA types were shown to be stable in chronically colonized patients and within outbreak-related strains, with excellent epidemiological concordance. The Burkholderia cepacia complex (BCC), which includes 18 closely related species, is mainly involved in pulmonary infections in patients with cystic fibrosis (CF), although it is also occasionally recovered in nosocomial infections and/or from immunocompromised hosts. BCC infection in CF patients is associated with accelerated decline of lung function, cepacia syndrome, and poor posttransplantation outcome. The most frequently recovered species of the BCC are B. cenocepacia and Burkholderia multivorans, which have been found to be responsible for large outbreaks among CF patients, prompting the implementation of infection control measures and epidemiological surveillance. B. cenocepacia consists of four recA subgroups: IIIA, which includes the Edinburgh-Toronto electrophoretic type 12 (ET-12) epidemic lineage (in Canada and the United Kingdom) (1); IIIB, which includes the PHDC epidemic lineage (in the United States) (2) and the Midwest clone (3); IIIC, which is exclusively environmental; and IIID, which was reported to be responsible for 50% of the BCC infections in an Italian CF center (4).Putative transmissibility markers have been identified within B. cenocepacia epidemic strains. The cblA pilin gene is characteristic of ET-12 strains (5). The B. cepacia epidemic strain marker (BCESM), which is part of a pathogenicity island, was identified in ET-12 strains and in other epidemic lineages but also in unique clinical or environmental strains (6, 7), whereas the PHDC lineage is BCESM negative. Finally, the IS1363 insertion sequence was demonstrated to be characteristic of the PHDC and ET...

show abstract

Section: Discussionmentioning

confidence: 99%

Development of a Multiple-Locus Variable-Number Tandem-Repeat Typing Scheme for Genetic Fingerprinting of Burkholderia cenocepacia and Application to Nationwide Epidemiological Analysis

Segonds¹,

Thouverez²,

Barthe³

et al. 2015

J Clin Microbiol

View full text Add to dashboard Cite

show abstract

“…The MLVA assay can provide detailed information for epidemiological monitoring of the chronic infections caused by P. aeruginosa. Thus, an automated MLVA assay was established to analyze 16 VNTRs using two multiplex PCRs with CE and was applied to genotype 83 isolates from eight patients with cystic fibrosis within 4 h. The results determined that most cases showed the same genotype during 8 years of chronic infection [31].…”

Section: Detecting and Diagnosing Bacteriamentioning

confidence: 99%

Capillary electrophoresis based on nucleic acid detection for diagnosing human infectious disease

Lian

Zhao

2016

Clinical Chemistry and Laboratory Medicine (CCLM)

View full text Add to dashboard Cite

Rapid transmission, high morbidity, and mortality are the features of human infectious diseases caused by microorganisms, such as bacteria, fungi, and viruses. These diseases may lead within a short period of time to great personal and property losses, especially in regions where sanitation is poor. Thus, rapid diagnoses are vital for the prevention and therapeutic intervention of human infectious diseases. Several conventional methods are often used to diagnose infectious diseases, e.g. methods based on cultures or morphology, or biochemical tests based on metabonomics. Although traditional methods are considered gold standards and are used most frequently, they are laborious, time consuming, and tedious and cannot meet the demand for rapid diagnoses. Disease diagnosis using capillary electrophoresis methods has the advantages of high efficiency, high throughput, and high speed, and coupled with the different nucleic acid detection strategies overcomes the drawbacks of traditional identification methods, precluding many types of false positive and negative results. Therefore, this review focuses on the application of capillary electrophoresis based on nucleic detection to the diagnosis of human infectious diseases, and offers an introduction to the limitations, advantages, and future developments of this approach.

show abstract

“…(NA) and STTR3 (211) and different combinations of the variable loci STTR5 (11,12,13) and STTR6 (9,10), where paranthesised numbers denote allele sizes and NA (no amplification) indicates negative PCR amplification, as previously described [2].…”

Section: Denmarkmentioning

confidence: 99%

“…The MLVA used for Mycobacterium tuberculosis [9] [10]. Alternatively, the assays can be set up locally, with no need to buy kits, since all the necessary information is published [10][11][12]. MLVA is also in routine use for Streptococcus pneumoniae, with more than 1,000 genotypes publicly accessible from the the Robert Picqué Military Hospital in Bordeaux [13] and for Acinetobacter baumanii [14].…”

Section: Francementioning

confidence: 99%

Use of multilocus variable-number tandem repeat analysis (MLVA) in eight European countries, 2012

et al. 2013

View full text Add to dashboard Cite

Genotyping of important medical or veterinary prokaryotes has become a very important tool during the last decades. Rapid development of fragment-separation and sequencing technologies has made many new genotyping strategies possible. Among these new methods is multilocus variable-number tandem repeat analysis (MLVA). Here we present an update on the use of MLVA in eight European countries (Denmark, France, Germany, Ireland, Italy, the Netherlands, Norway and Sweden). Researchers in Europe have been active in developing and implementing a large array of different assays. MLVA has been used as a typing tool in several contexts, from aiding in resolving outbreaks of food-borne bacteria to typing organisms that may pose a bioterrorist threat, as well as in scientific studies.

show abstract

A new highly discriminatory multiplex capillary-based MLVA assay as a tool for the epidemiological survey of Pseudomonas aeruginosa in cystic fibrosis patients

Cited by 32 publications

References 29 publications

Development of a Multiple-Locus Variable-Number Tandem-Repeat Typing Scheme for Genetic Fingerprinting of Burkholderia cenocepacia and Application to Nationwide Epidemiological Analysis

Development of a Multiple-Locus Variable-Number Tandem-Repeat Typing Scheme for Genetic Fingerprinting of Burkholderia cenocepacia and Application to Nationwide Epidemiological Analysis

Capillary electrophoresis based on nucleic acid detection for diagnosing human infectious disease

Use of multilocus variable-number tandem repeat analysis (MLVA) in eight European countries, 2012

Contact Info

Product

Resources

About