Identification of
            <i>Staphylococcus</i>
            spp. by PCR-Restriction Fragment Length Polymorphism of
            <i>gap</i>
            Gene

Yugueros, J; Temprano, A; Sánchez, María de la Paz; Luengo, José M.; Naharro, Germán

doi:10.1128/jcm.39.10.3693-3695.2001

Cited by 89 publications

(38 citation statements)

References 39 publications

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“…Strains were identified by Gram staining, catalase test, coagulase test in rabbit plasma (Remel), agglutination test and API20Staph Gallery (bioMerieux). Then, identification was completed by a PCR-restriction fragment length polymorphism using the 'gap' gene as previously recommended by Yugueros et al (2001) and Karakulska et al (2012).…”

Section: Methodsmentioning

confidence: 99%

Staphylococcal cassette chromosome mec typing and mecA sequencing in methicillin-resistant staphylococci from Algeria: a highly diversified element with new mutations in mecA

Djoudi

Bonura

Touati

et al. 2016

Journal of Medical Microbiology

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Genetic mechanisms of methicillin resistance are still relevant in staphylococci. The aims of this study are to assess the possible exchanges of staphylococcal cassette chromosome mec (SCCmec) among isolates of methicillin-resistant staphylococci (MRS) and to check for known or new mutations in mecA DNA. A total of 35 MRS non-repetitive isolates were recovered, including 20 Staphylococcus haemolyticus, 7 Staphylococcus aureus, 4 Staphylococcus sciuri, 2 Staphylococcus saprophyticus and 1 isolate each of Staphylococcus xylosus and Staphylococcus lentus. Only 16 of the 35 strains were assigned to known SCCmec types: 7 SCCmec VII, 6 SCCmec IV and 3 SCCmec III, with possible horizontal transfer of the SCCmec VII from methicillin-resistant S. haemolyticus to methicillin-susceptible S. aureus. mecA gene sequencing in ten selected isolates allowed description of nine punctual mutations, seven of which were reported for the first time. The most frequent mutation was G246E, identified in isolates of methicillin-resistant S. aureus, S. sciuri, S. saprophyticus and S. lentus. These results emphasized the high degree of genetic diversity of SCCmec element in MRS and describe new missense mutations in mecA, which might be important in understanding the evolution of methicillin and new b-lactam resistance.

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Section: Methodsmentioning

confidence: 99%

Staphylococcal cassette chromosome mec typing and mecA sequencing in methicillin-resistant staphylococci from Algeria: a highly diversified element with new mutations in mecA

Djoudi

Bonura

Touati

et al. 2016

Journal of Medical Microbiology

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“…PCR-RFLP of gap gene: The genetic identification of the strains was achieved by carrying out a PCR-RFLP analysis by amplifying the gap gene, encoding glyceraldehyde-3-phosphate dehydrogenase, as previously described by Yugueros et al 2001(Yugueros et al 2001). The obtained PCR product (933 bp) was digested with the restriction enzyme AluI (Thermo Fisher Scientific, Waltham, USA) and separated by electrophoresis gel.…”

Section: Molecular Methodsmentioning

confidence: 99%

Clustering of Staphylococcus aureus bovine mastitis strains from regions of Central-Eastern Poland based on their biochemical and genetic characteristics

Puacz¹,

Ilczyszyn²,

Kosecka³

et al. 2015

Polish Journal of Veterinary Sciences

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Staphylococcus aureus strains were isolated from mastitic milk of cows with infected mammary glands. The animals were living in 12 different farms near Lublin, in Central-Eastern Poland. A biochemical identification method based on enzymatic assay was performed, followed by haemolytic and proteolytic tests. PCR-RFLP targeted on the gap gene allowed the genetic identification of strains at the species level and verified phenotypic identification results. A molecular typing method using triplex PCR was performed to recognize the genetic similarity of the analyzed strains. DNA microarray hybridization (StaphyType, Alere Technologies) was used for detection of antibiotic resistance and virulence associated markers. The results obtained indicate high genetic similarity in strains isolated from the same sites. High genetic similarities were also detected between strains isolated from cows from different farms of the same region. A slightly lower similarity was noted however, in strains from various regions indicating that the strains are herd specific and that the cow's infections caused by S. aureus were of a clonal character. In 21 representative isolates selected for DNA-microarray testing, only fosfomycin (fosB) and penicillin resistance markers (blaZ, blaI, blaR) were detected. The presence of genes coding for haemolysins

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“…The specific primer sequences for Staphylococcus aureus and Staphylococcus epidermidis have been previously described 7 . In addition, we used a generic staphylococcal primer set (GF-1/GR-2) directed against the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene 16 and a 23S rRNA Staphylococcus aureus-specific primer set (532720-GGACGA-CATTAGACGAATCA-532739 and 534038-CGGGCACCTATT-TTCTATCT-534019), which were developed and validated in the present study under the same polymerase-chain-reactioncycling conditions as previously described 7 .…”

Section: Confocal Laser Scanning Microscopy and Viability Stainingmentioning

confidence: 99%

Direct Demonstration of Viable Staphylococcus aureus Biofilms in an Infected Total Joint Arthroplasty

Stoodley

Nistico

Johnson

et al. 2008

The Journal of Bone and Joint Surgery-American Volume

160

129

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Identification of Staphylococcus spp. by PCR-Restriction Fragment Length Polymorphism of gap Gene

Cited by 89 publications

References 39 publications

Staphylococcal cassette chromosome mec typing and mecA sequencing in methicillin-resistant staphylococci from Algeria: a highly diversified element with new mutations in mecA

Staphylococcal cassette chromosome mec typing and mecA sequencing in methicillin-resistant staphylococci from Algeria: a highly diversified element with new mutations in mecA

Clustering of Staphylococcus aureus bovine mastitis strains from regions of Central-Eastern Poland based on their biochemical and genetic characteristics

Direct Demonstration of Viable Staphylococcus aureus Biofilms in an Infected Total Joint Arthroplasty

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