Rapid assay for detection of methicillin-resistantStaphylococcus aureususing multiplex PCR

Barski, Piotr; Piechowicz, Lidia; Galiński, J; Kur, Józef

doi:10.1006/mcpr.1996.0066

Cited by 74 publications

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“…As methicillin resistance mediated by PBP 2a is often heterogeneously expressed in staphylococci (2,7,24), PCR detection of mecA gene is the "gold standard" for the detection of methicillin resistance (1,2,10,16,21). In the case of severe infections, it is clinically useful to provide rapid identification and antimicrobial susceptibility test results (5).…”

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Evaluation of a Triplex PCR Assay To DiscriminateStaphylococcus aureusfrom Coagulase-Negative Staphylococci and Determine Methicillin Resistance from Blood Cultures

et al. 2002

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A triplex PCR targeting the 16S rRNA, mecA, and nuc genes was developed for identification of staphylococci and detection of methicillin resistance. After validation of the assay with a collection of strains of staphylococci and enterococci (n ‫؍‬ 169), the assay was evaluated with cultures of blood with gram-positive cocci from 40 patients. Accurate results were obtained for 59 (98%) of 61 cultures within 6 h of growth detection.

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Evaluation of a Triplex PCR Assay To DiscriminateStaphylococcus aureusfrom Coagulase-Negative Staphylococci and Determine Methicillin Resistance from Blood Cultures

et al. 2002

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“…The correct identification of S. aureus and the detection of the mecA gene based on molecular methods have evolved as the method of choice for definitive identification. Earlier studies, however, involved more or less complicated manual DNA extraction protocols, followed by single or multiplex PCR with detection of ampli- fication products on agarose gels (1,4,11,13,15,23,24,28,36). Those assays, however, were time-consuming, prone to contamination, and not suitable for routine diagnostic laboratories because of the lack of a hybridization technique.…”

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“…Detection of the mecA gene by PCR has been described as a rapid method for the identification of MRSA (1,4,11,13,15,23,24,28,36).…”

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Detection of Methicillin-Resistant Staphylococcus aureus and Simultaneous Confirmation by Automated Nucleic Acid Extraction and Real-Time PCR

et al. 2002

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A molecular assay for the simultaneous detection of a Staphylococcus aureus-specific gene and the mecA gene, responsible for the resistance to methicillin in staphylococci, was evaluated. The assay included an automated DNA extraction protocol conducted with a MagNA Pure instrument and real-time PCR conducted with a LightCycler instrument. The performance and robustness of the assay were evaluated for a suspension of methicillin-resistant S. aureus (MRSA) strain with a turbidity equivalent to a McFarland standard of 0.5, which was found to be the ideal working concentration. The specificity of the new molecular assay was tested with a panel of 30 gram-negative and gram-positive bacterial strains other than MRSA. No cross-reactivity was observed. In a clinical study, 109 isolates of MRSA were investigated. All clinical MRSA isolates gave positive results for the S. aureus-specific genomic target, and all but one were positive for the mecA gene. In conclusion, the new molecular assay was found to be quick, robust, and laborsaving, and it proved to be suitable for a routine molecular diagnostic laboratory.

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“…Bu yüzden polimeraz zincir reaksiyonu (PCR) esaslı moleküler yöntemler antibiyotik direnç genlerinin belirlenmesinde sıklıkla tercih edilmektedir [8] . …”

Section: Phenotypic and Genotypic Determination Of Antibiotic Resistaunclassified

Phenotypic and Genotypic Determination of Antibiotic Resistances of Some Clinical Staphylococcus aureus Isolates

Orhan¹,

Kayiş²,

Akyol³

et al. 2017

FLORA

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ÖZET Giriş: Staphylococcus aureus, ciddi hastalıklara yol açabilen tehlikeli bir patojendir. Antibiyotik direncinin ortaya çıkması büyük oranda etkili tedaviler bulma sorununa neden olmuştur. Bu çalışma, hastaların çeşitli klinik örneklerinden elde edilen S. aureus izolatlarında bazı antibiyotiklerin antibiyotik duyarlılık paternleri ile antibiyotik direnç genleri arasındaki ilişkiyi değerlendirmek amacıyla yapılmıştır. Materyal ve Metod:Toplam 100 S. aureus klinik izolatı, antimikrobik duyarlılık testine tabi tutulmuştur. Oksasilin (mecA), trimetoprim-sülfametoksazol (TMP-SMZ) (dfrA), eritromisin, klindamisin (ermA, ermB, ermC ve msrA), kinupristin-dalfopristin (vatA, vatB, vatC) ve siprofloksasin (gyrA, gyrB) GİRİŞStaphylococcus aureus nispeten önemsiz yüzey-sel deri infeksiyonlarından derin doku infeksiyonu ve bakteremi arasında değişen, çeşitli insan infeksiyonları arasında tıbbi önemi olan bir patojendir. S. aureus'un özellikle antibiyotiğe dirençli izolatları, ağırlıklı olarak ağır sepsis ve septik şok ile sonuçlanan hastane infeksiyonlarında tedavi güçlü-ğüne yol açarak yüksek morbidite ve mortaliteye neden olmaktadır. Bu durum da dünya çapında sağlık üzerinde önemli bir yük oluşturmaktadır [1] .Stafilokoklarda metisilin direnci, mecA geni tarafından kodlanan ve penisilin bağlayan protein 2a (PBP2a) olarak adlandırılan bir proteinin üretimi sonucu ortaya çıkar. Bu modifiye olmuş proteinin beta-laktam grubu antibiyotiklere afinitesi düşük olduğu için tüm beta-laktam ve beta-laktam türevi antibiyotiklere dirence neden olmaktadır [2] .Stafilokoklarda trimetoprime iki farklı direnç mekanizması tanımlanmıştır: biri kromozomal dfrA geninde nokta mutasyonlar, diğeri ise dihidrofolat redüktaz (DHFR)'ın ilaca dirençli varyantları-nı kodlayan tipik olarak plazmid kaynaklı direnç belirleyicilerinin yatay olarak edinimidir. S. aureus'daki endojen DHFR mutasyonu, düşük-orta seviyede direnç kazandırırken, yatay olarak edinilmiş DHFR'nin ekspresyonu yüksek seviyeli dirence yol açar [3] .S. aureus'da makrolidler, linkozamidler ve streptogramin B (MLS B ) antibiyotiklerine karşı çapraz direnç, eritromisin ribozom metilasyon (erm) genleri tarafından kodlanır. Bu tip direnç konstitütif veya indüklenebilir olabilir. MLS B indüklenebilir fenotipi eritromisine dirençli S. aureus'da klindamisin direncine neden olabilir. Böylece klindamisin tedavisinin klinik başarısızlığına yol açar [4] .

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Rapid assay for detection of methicillin-resistantStaphylococcus aureususing multiplex PCR

Cited by 74 publications

References 0 publications

Evaluation of a Triplex PCR Assay To DiscriminateStaphylococcus aureusfrom Coagulase-Negative Staphylococci and Determine Methicillin Resistance from Blood Cultures

Evaluation of a Triplex PCR Assay To DiscriminateStaphylococcus aureusfrom Coagulase-Negative Staphylococci and Determine Methicillin Resistance from Blood Cultures

Detection of Methicillin-Resistant Staphylococcus aureus and Simultaneous Confirmation by Automated Nucleic Acid Extraction and Real-Time PCR

Phenotypic and Genotypic Determination of Antibiotic Resistances of Some Clinical Staphylococcus aureus Isolates

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