Journal: FEMS microbiology letters
Article Title: Development and evaluation of hexaplex PCR for rapid detection of methicillin, cadmium/zinc and antiseptic-resistant staphylococci, with simultaneous identification of PVL-positive and -negative Staphylococcus aureus and coagulase negative staphylococci.
doi: 10.1111/1574-6968.12383
Figure Lengend Snippet: Fig. 1. (a) Agarose gel electrophoresis of simplex products obtained with reference strains of Staphylococcus aureus and Staphylococcus epiderimidis and stained with ethidium bromide. Lane M, 100-bp DNA ladder (NEB); lane 1, S. aureus ATCC 43300 (16S rRNA+); lane 2, Staphylococcus epidermidis ATCC 12228 (czrC+); lane 3, S. aureus ATCC 25923 (pvl+); lane 4, S. aureus ATCC 43300 (nuc+); Lane 5, S. aureus ATCC 43300 (mecA+); lane 6, S. aureus Mu50 (qacA/B+). (b) Agarose gel electrophoresis of hexaplex PCR products obtained with reference and control strains of S. aureus and CoNS and stained with ethidium bromide. Lane M, 100-bp DNA ladder (NEB); lane 1, S. aureus ATCC 43300 (16S rRNA+, nuc+, mecA+); lane 2, S. aureus Mu50 (16S rRNA+, nuc+, mecA+, qacA/B+); lane 3, S. aureus ATCC 25923 (16S rRNA+, nuc+, pvl+); lane 4, S. aureus 1295 (16S rRNA+, nuc+, pvl+, mecA+); lane 5, S. epidermidis ATCC 12228 (16S rRNA+, czrC+); lane 6, S. epidermidis 1056 (16S rRNA+, mecA+, qacA/B+); lane 7, Staphylococcus haemolyticus ATCC 29970 (16S rRNA+); lane 8, S. haemolyticus 119 (16S rRNA+, mecA+, czrC+, qacA/B+); lane 9, S. haemolyticus 263 (16S rRNA+, mecA+, czrC+); lane 10, Staphylococcus saprophyticus ATCC 15305 (16S rRNA+); lane 11, Staphylococcus simulans ATCC 27851 (16S rRNA+, czrC+); lane 12, Staphylococcus warneri ATCC 49454 (16S rRNA+); lane 13, Staphylococcus gallinarum ATCC 700401 (16S rRNA+); lane 14, Staphylococcus sciuri ATCC 29060 (16S rRNA+); lane 15, Staphylococcus xylosus ATCC 29971 (16S rRNA+); lane 16, Staphylococcus lugdunensis ATCC 49576 (16S rRNA+); lane 17, Staphylococcus lentus ATCC 700403 (16S rRNA+); lane 18, Staphylococcus capitis 527 (16S rRNA+, mecA+); lane 19, Streptococcus pyogenes ATCC 19615; lane 20, Streptococcus pneumoniae and lane 21, Milli-Q.
Article Snippet: The DNA was fixed to the nylon membrane using UV cross-linker (Bio-Rad) for 2 min. PCR amplified products obtained from S. aureus strain Mu50 for staphylococcal 16S rRNA, nuc, mecA, qacA/B genes; S. aureus strain ATCC 25923 for pvl gene and S. epidermidis strain ATCC 12228 for czrC gene were used as probes, randomly labeled (Feinberg & Vogelstein, 1984) with [a-32P] dCTP (3000 Ci mmol 1; BARC, Bombay, India) and hybridized at 65 °C in phosphate buffer containing 500 mM Na2HPO4 (pH 7.2), 7% (w/v) sodium dodecyl sulfate (SDS), 1 mM EDTA, and 1% (w/ v) bovine serine albumin.
Techniques: Agarose Gel Electrophoresis, Staining, Control
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