A Universal Approach To Bacterial Molecular Epidemiology By Polymerase Chain Reaction Ribotyping

Kostman, Jay R.; Alden, Mitchell B.; Mair, Michael; Edlind, Thomas D.; LiPuma, John J.; Stull, Terrence L.

doi:10.1093/infdis/171.1.204

Cited by 97 publications

(61 citation statements)

References 12 publications

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“…The ribotyping-polymerase chain reaction (PCR) is often referenced and used because of its high taxonomic and epidemiological value. This technique is a valuable tool for identifying and differentiating isolates of the Staphylococcus genus [13][14][15] .…”

Section: Methodsmentioning

confidence: 99%

The occurrence and dissemination of methicillin and vancomycin-resistant Staphylococcus in samples from patients and health professionals of a university hospital in Recife, State of Pernambuco, Brazil

Rabelo

Neto

Loibman

et al. 2014

Rev. Soc. Bras. Med. Trop.

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Introduction: Methicillin-resistant Staphylococcus aureus (MRSA) strains have been responsible for many nosocomial outbreaks. Within hospitals, colonized employees often act as reservoirs for the spread of this organism. This study collected clinical samples of 91 patients admitted to the intensive care unit (ICU), hemodialysis/nephrology service and surgical clinic, and biological samples from the nasal cavities of 120 professionals working in those environments, of a University Hospital in Recife, in the State of Pernambuco, Brazil. The main objective of this study was to determine the occurrence and dissemination of methicillin-and vancomycin-resistant Staphylococcus spp. Methods: The isolates obtained were tested for susceptibility to oxacillin and vancomycin and detection of the mecA gene. In addition, the isolates were evaluated for the presence of clones by ribotyping-polymerase chain reaction (PCR). Results: MRSA occurrence, as detected by the presence of the mecA gene, was more prevalent among nursing technicians; 48.1% (13/27) and 40.7% (11/27) of the isolates were from health professionals of the surgical clinic. In patients, the most frequent occurrence of mecA-positive isolates was among the samples from catheter tips (33.3%; 3/9), obtained mostly from the hemodialysis/nephrology service. Eight vancomycin-resistant strains were found among the MRSA isolates through vancomycin screening. Based on the amplifi cation patterns, 17 ribotypes were identifi ed, with some distributed between patients and professionals. Conclusions: Despite the great diversity of clones, which makes it diffi cult to trace the source of the infection, knowledge of the molecular and phenotypic profi les of Staphylococcus samples can contribute towards guiding therapeutic approaches in the treatment and control of nosocomial infections.

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Section: Methodsmentioning

confidence: 99%

The occurrence and dissemination of methicillin and vancomycin-resistant Staphylococcus in samples from patients and health professionals of a university hospital in Recife, State of Pernambuco, Brazil

Rabelo

Neto

Loibman

et al. 2014

Rev. Soc. Bras. Med. Trop.

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“…However, the lack of discriminatory power of such techniques has led researchers to develop alternative molecular-based methods (10,11).…”

Section: Introductionmentioning

confidence: 99%

Typing of Enterococcus faecium by polymerase chain reaction and pulsed field gel electrophoresis

Bedendo

Pignatari

2000

Braz J Med Biol Res

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Polymerase chain reaction (PCR) with JB1 or REP consensus oligonucleotides and pulsed field gel electrophoresis (PFGE) were used to study genomic DNA extracted from 31 strains of enterococci. Eleven ATCC strains, representative of 11 species of Enterococcus, were initially tested by JB1-PCR, revealing that Enterococcus malodoratus and Enterococcus hirae presented identical banding patterns. Eight Enterococcus faecium isolates from Stanford University and 12 from São Paulo Hospital were studied by JB1-PCR, REP-PCR 1/2R and PFGE. Among the isolates from Stanford University, 5 genotypes were defined by JB1-PCR, 7 by REP-PCR 1/2R and 4 by PFGE. Among the isolates from São Paulo Hospital, 9 genotypes were identified by JB1-PCR, 6 by REP-PCR and 5 by PFGE. The three methods identified identical genotypes, but there was not complete agreement among them.

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“…Contrary to expected species consistency, roughly the same level of variation was observed within species as between them. Part of this problem derives from the organization of rRNA genes (rDNA) as a multigene family, often as rrn operons that, in bacteria, are present in numbers varying from 1 to 11 (Gurtler, 1993 ;Hill & Harnish, 198 1 ;Kostman et al, 1992;Suzuki & Yamada, 1988) and a few hundred copies in eukaryotic genomes (Long & David, 1980). The members of a multigene family are subjected to a homogenization process (Dover, 1982(Dover, , 1987Ohta, 199 1) by molecular interaction mechanisms, such as gene conversion, and rDNA sequences tend to evolve in concert.…”

Section: Introductionmentioning

confidence: 99%

Patterns of sequence variation in two regions of the 16S rRNA multigene family of Escherichia coli

Martínez‐Murcia¹,

Antón²,

Rodrı́guez-Valera³

1999

International Journal of Systematic and Evolutionary Microbiology

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Sequence heterogeneities of variable positions located a t regions V1 and V6 of 56 cloned 16s rRNA genes were determined from six Escherichia coli strains. These nucleotides were involved in secondary structure base-pairing of stem-loops. Compensatory and single mutations have occurred but secondary structure was conserved. Eight different sequences were found in the stem a t region V1 indicating that in these sites mutation rates are higher than those of homogenization processes. Region V6 showed two different structures (V6-I and V6-ll) although heterogeneities were determined in nine sites. Strains ECOR52 and ECOR56 only showed the V6-I sequence, ECOR35 showed V6-ll, whereas clones from ECOR42 and ECOR49 showed both types of V6 structures. Results were confirmed b y PCR using V6 sequence-specific probes. Stem V6-ll was also found in 165 rRNA sequences deposited in the RDP (Ribosomal Database Project) belonging to distantly related taxa; ancestral sequence V6-II seems to be homogenized in all rm operons of the multigene family of strain ECOR35 producing effects of distortion in the molecular clock, similar to those that homoplasies could produce. V6 sequence-specif ic probes were applied to the 72 ECOR strains: half showed both V6-I and V6-ll, and the rest had one or another. Only strain ECOR24 did not yield products in the PCR test and sequencing of 12 cloned 165 rRNA genes revealed a third form, V6-Ill, also found in the RDP. Concerted evolution by homogenization of the rRNA family may induce chronometric distortions responsible for a loss of ultrametricity in phylogenetic trees, particularly, of very closely related micro-organisms.

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A Universal Approach To Bacterial Molecular Epidemiology By Polymerase Chain Reaction Ribotyping

Cited by 97 publications

References 12 publications

The occurrence and dissemination of methicillin and vancomycin-resistant Staphylococcus in samples from patients and health professionals of a university hospital in Recife, State of Pernambuco, Brazil

The occurrence and dissemination of methicillin and vancomycin-resistant Staphylococcus in samples from patients and health professionals of a university hospital in Recife, State of Pernambuco, Brazil

Typing of Enterococcus faecium by polymerase chain reaction and pulsed field gel electrophoresis

Patterns of sequence variation in two regions of the 16S rRNA multigene family of Escherichia coli

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