2003
DOI: 10.3346/jkms.2003.18.5.631
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Multiplex PCR for the Detection of Genes Encoding Aminoglycoside Modifying Enzymes and Methicillin Resistance among Staphylococcus Species

Abstract: We developed multiplex polymerase chain reaction (PCR) to detect aac(6 ')/aph(2 "), aph(3 ')-IIIa, and ant(4 ')-Ia, the genes encoding the most clinically relevant amino-glycoside modifying enzymes (AME), and simultaneously, the methicillin resistant gene, mecA, in Staphylococcus species. Clinical isolates of 45 S. aureus and 47 coagulase negative staphylococci (CNS) from tertiary university hospitals were tested by conventional susceptibility testing, using the agar dilution method and by multiplex PCR. Of a … Show more

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Cited by 120 publications
(85 citation statements)
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“…All isolates were tested for resistance to aminoglycosides by a disk diffusion method using amikacin 30 μg, gentamicin 10 μg, tobramycin 10 μg and kanamycin 30 μg, 6 according to the Clinical and Laboratory Standards Institute guidelines [3].…”
Section: Susceptibility Testingmentioning
confidence: 99%
“…All isolates were tested for resistance to aminoglycosides by a disk diffusion method using amikacin 30 μg, gentamicin 10 μg, tobramycin 10 μg and kanamycin 30 μg, 6 according to the Clinical and Laboratory Standards Institute guidelines [3].…”
Section: Susceptibility Testingmentioning
confidence: 99%
“…mecA geni heterojen olarak eksprese edilebildiği için tüm metisiline dirençli stafilokok suşları fenotipik yön-temlerle saptanmayabilmektedir. Bu yüzden mecA geninin saptanması için kullanılan PCR tekniği konvansiyonel yöntemlerle karşılaştırıldığında hem daha hızlı hem de güvenilir olan bir altın standart yöntem olarak kabul edilmektedir [14] . Çalışmamızda metisilin disk difüzyon ve Phoenix yöntemleriyle test edildiğinde metisiline direnç %19 idi ve bu izolatların hepsi de mecA genine sahipti.…”
Section: Discussionunclassified
“…Bizim çalışmamızın aksine birçok çalışma raporunda en sık genetik belirleyici olarak ermA geni tespit edilmiştir [4,10,[12][13][14][15][16][17][18][19][20][21][22] .…”
Section: Discussionunclassified
“…All CoNS were also screened by PCR for the presence of the mecA gene, as previously reported (1). The primers used to detect mecA gene were: 5′-CCTAG TAAAGCTCCGGAA-3′ and 5′-CTAGTCCATTC GGTCCA-3′, and the PCR reaction was as follows: initial denaturation at 95°C for 2 min, denaturation at 94°C for 2 min (30 cycles), annealing at 58°C for 30 sec and extension at 72°C for 30 sec, with a final extension step at 72°C for 7 min.…”
Section: Methodsmentioning
confidence: 99%