Triplex real-time polymerase chain reaction assay for simultaneous detection of Staphylococcus aureus and coagulase-negative staphylococci and determination of methicillin resistance directly from positive blood culture bottles

Kılıç, Abdullah; Muldrew, Kenneth L.; Tang, Yi‐Wei; Başustaoğlu, Ahmet

doi:10.1016/j.diagmicrobio.2009.11.010

Cited by 51 publications

(39 citation statements)

References 41 publications

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“…Both of the latter PCR assays can be used on positive blood culture vials that show Gram-positive cocci in clusters by Gram stain. In addition, a recently published evaluation of an in-housedeveloped triplex PCR assay for the simultaneous detection of S. aureus and coagulase-negative staphylococci and determination of meticillin resistance directly from blood culture bottles reported sensitivities of 99.7%, 100% and 99.2% and specificities of 100%, 100% and 98.7%, respectively, for the tuf, nuc and mecA gene targets [65].…”

Section: Clinical Diagnosis Of Mrsamentioning

confidence: 99%

Update on screening and clinical diagnosis of meticillin-resistant Staphylococcus aureus (MRSA)

Harbarth

Hawkey

Tenover³

et al. 2011

International Journal of Antimicrobial Agents

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Section: Clinical Diagnosis Of Mrsamentioning

confidence: 99%

Update on screening and clinical diagnosis of meticillin-resistant Staphylococcus aureus (MRSA)

Harbarth

Hawkey

Tenover³

et al. 2011

International Journal of Antimicrobial Agents

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“…mecA gen varlığı, gerçek zamanlı polimeraz zincir reaksiyonu (PCR) ile Kılıç 4,[20][21][22][23] . Çalışmamızda, ülkemizdeki çalışmalara benzer olarak, 523 MRSA izolatının dokuzu (%1.7) ve 1177 MSSA izolatının 23 (%1.9)'ü PVL pozitif olarak bulunmuştur.…”

Section: Pvl Meca Ve Mecc Genlerinin Saptanmasıunclassified

Investigation of the Presence of mecC and Panton-Valentine Leukocidin Genes in Staphylococcus aureus Strains Isolated from Clinical Specimens During Seven Years Period

Kılıç

Dogan²,

Kaya³

et al. 2015

Mikrobiyol Bul

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ÖZKlinik mikrobiyoloji laboratuvarlarında metisiline dirençli Staphylococcus aureus (MRSA)'un saptanması ve tanımlanması, uygun tedavi seçimi ve epidemiyolojik verilerin elde edilmesi için önemlidir. mecC geni, mecA genine yaklaşık %69 DNA benzerliği gösteren bir mecA homoloğudur ve PBP2a/2' proteinine yaklaşık %63 benzerlik gösteren proteini kodlamaktadır. Çeşitli çalışmalar, mecC pozitif MRSA izolatlarının, çapraz kontaminasyon ile çiftlik hayvanlarından insanlara bulaştığını desteklemektedir. Panton-Valentine lökosidini (PVL) ise, S.aureus'un potent sitotoksini olup, önemli bir virülans faktörü olarak kabul edilmektedir. Bu çalışmada, klinik örneklerden izole edilen S.aureus suşlarında mecC ve pvl gen varlığının ve prevalansının araştırılması amaçlanmıştır. Çalışmaya, Ocak 2007-Aralık 2014 tarihleri arasında hastanemizde izole edilmiş olan 1177'si metisiline duyarlı S.aureus (MSSA) ve 523'ü MRSA olmak üzere toplam 1700 S.aureus izolatı dahil edilmiştir. İzolatlar, konvansiyonel yöntemler ve BD Phoenix otomatize sistemi (BD Diagnostic Instrument Systems, ABD) ile tanımlanmıştır. Antibiyotik duyarlılık testi, oksasilin (1 μg) ve sefoksitin (30 μg) için Kirby-Bauer disk difüzyon yöntemiyle CLSI standartlarına göre yapılmıştır. mecA gen varlığı gerçek zamanlı PCR; pvl ve mecC gen varlığı ise konvansiyonel PCR yöntemi ile araştırılmıştır. Suşların izole edildiği hastaların %44.6 (759/1700)'sı poliklinik hastası ve %65.8 (1119/1700)'i erkek olup, yaş ortalaması 39.7 yıldır. Çalışmamızda değerlendirilen 1700 izolatın 523 (%30.7)'ünde mecA geni pozitif bulunmuş; hiçbir izolatta mecC geni tespit edilmemiştir. MSSA izolatlarının %1.9 (23/1177)'u ve MRSA izolatlarının %1.7 (9/523)'sinde olmak üzere, toplam 32 (%1.8) izolatta pvl geni pozitif bulunmuş-tur. PVL pozitif S.aureus'ların %56.2 (18/32)'sinin, deri ve yumuşak doku enfeksiyonu örneklerinden izole Geliş Tarihi

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“…PCR primers and probes (Table 2) were custom synthesized by Sigma-Aldrich. For S. aureus, a 96 bp fragment of the nuc gene was amplified and detected (Kilic et al, 2010), whilst for GAS, a 154 bp fragment of the ntpC gene was amplified and detected (Hung et al, 2012). PCR of a T4 bacteriophage internal process control confirmed successful DNA and inhibitor-free nucleic acid extracts.…”

Section: Methodsmentioning

confidence: 85%

Specific PCR, bacterial culture, serology and pharyngeal sampling to enhance the aetiological diagnosis of cellulitis

Toleman

Vipond

Brindle

2016

Journal of Medical Microbiology

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It is often difficult to obtain a bacteriological diagnosis in patients with cellulitis. We examined the utility of molecular techniques and skin and throat cultures, as well as serology, in providing evidence of either Staphylococcus aureus or group A Streptococcus (GAS) presence in patients with cellulitis. Samples were collected from patients with a clinical diagnosis of cellulitis who were recruited into a prospective placebo-controlled clinical trial (C4C study, EudraCT 2013-001218-14). Specific PCR, paired serology and culture for both organisms were carried out on a variety of samples where appropriate. Despite utilizing a range of diagnostic methods, a bacteriological diagnosis was only achieved in 43 % of patients with a clinical diagnosis of cellulitis. Seventeen per cent of patients tested positive for GAS by any method but only 4 % were positive by PCR, whilst S. aureus was detected in 34% of samples. Bacterial diagnosis in cases of cellulitis remains challenging. This is probably due to a very low bacterial burden with toxin production resulting in inflammation mediating skin damage. Further consideration for the need for long courses of antimicrobial therapy for cellulitis therefore appears merited.

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Triplex real-time polymerase chain reaction assay for simultaneous detection of Staphylococcus aureus and coagulase-negative staphylococci and determination of methicillin resistance directly from positive blood culture bottles

Cited by 51 publications

References 41 publications

Update on screening and clinical diagnosis of meticillin-resistant Staphylococcus aureus (MRSA)

Update on screening and clinical diagnosis of meticillin-resistant Staphylococcus aureus (MRSA)

Investigation of the Presence of mecC and Panton-Valentine Leukocidin Genes in Staphylococcus aureus Strains Isolated from Clinical Specimens During Seven Years Period

Specific PCR, bacterial culture, serology and pharyngeal sampling to enhance the aetiological diagnosis of cellulitis

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