dc.contributor.author | İstanbullu Tosun, Ayşe | |
dc.contributor.author | Kolukırık, Mustafa | |
dc.contributor.author | Yılmaz, Mesut | |
dc.contributor.author | Nar Ötgün, Selin | |
dc.contributor.author | Aygün, Gökhan | |
dc.contributor.author | Ketre Kolukırık, Canan Zöhre | |
dc.contributor.author | Zeybek, Ümit | |
dc.contributor.author | Girgin Özgümüş, Gözde | |
dc.contributor.author | Turan, Meral | |
dc.contributor.author | Kuşkucu, Mert | |
dc.contributor.author | İnce, Orhan | |
dc.contributor.author | İnce, Bahar | |
dc.contributor.author | Kılıç, Selçuk | |
dc.date.accessioned | 2023-03-03T09:50:56Z | |
dc.date.available | 2023-03-03T09:50:56Z | |
dc.date.issued | 2023 | en_US |
dc.identifier.citation | İstanbullu Tosun, A., Kolukırık, M., Yılmaz, M., Nar Ötgün, S., Aygün, G., Ketre Kolukırık, C. Z. ... Kılıç, S. (2023). Development of a new multiplex real-time PCR assay for rapid screening of hospital-acquired infection agents. Journal of Microbiological Methods, 206. https://dx.doi.org/10.1016/j.mimet.2023.106690 | en_US |
dc.identifier.issn | 0167-7012 | |
dc.identifier.issn | 1872-8359 | |
dc.identifier.uri | https://dx.doi.org/10.1016/j.mimet.2023.106690 | |
dc.identifier.uri | https://hdl.handle.net/20.500.12511/10563 | |
dc.description.abstract | Aims: A new multiplex real-time PCR (qPCR) assay was developed to detect antibiotic-resistant hospital-acquired infectious agents in nasal and rectal swab samples in 1.5 h without the need for nucleic acid extraction. Methods: Spiked negative clinical specimens were used for the analytical performance evaluation. Double-blind samples were collected from 1788 patients to assess the relative clinical performance of the qPCR assay to the conventional culture-based methods. Bio-Speedy® Fast Lysis Buffer (FLB) and 2× qPCR-Mix for hydrolysis probes (Bioeksen R&D Technologies, Istanbul, Turkey) and LightCycler® 96 Instrument (Roche Inc., Branchburg, NJ, USA) were used for all molecular analyses. The samples were transferred into 400 L FLB, homogenized and immediately used in qPCRs. The target DNA regions are vanA and vanB genes for vancomycin-resistant Enterococcus (VRE); blaKPC, blaNDM, blaVIM, blaIMP, blaOXA-23, blaOXA-48, blaOXA-58 genes for carbapenem-resistant Enterobacteriaceae (CRE); and mecA, mecC and spa for methicillin-resistant Staphylococcus aureus (MRSA). Results: No qPCR tests produced positive results for the samples spiked with the potential cross-reacting organisms. The limit of detection (LOD) of the assay for all targets was 100 colony-forming unit (cfu)/swab-sample. Results of the repeatability studies in two different centers were in 96%–100% (69/72–72/72) agreement. The relative specificity and sensitivity of the qPCR assay were respectively 96.8% and 98.8% for VRE; 94.9% and 95.1% for CRE; 99.9% and 97.1% for MRSA. Conclusions: The developed qPCR assay can screen antibiotic-resistant hospital-acquired infectious agents in infected/colonized patients with an equal clinical performance to the culture-based methods. | en_US |
dc.language.iso | eng | en_US |
dc.publisher | Elsevier B.V. | en_US |
dc.rights | info:eu-repo/semantics/embargoedAccess | en_US |
dc.subject | Carbapenem-Resistant Enterobacteriaceae | en_US |
dc.subject | Hospital-Acquired Infection | en_US |
dc.subject | Methicillin-Resistant Staphylococcus Aureus | en_US |
dc.subject | Real-Time PCR | en_US |
dc.subject | Vancomycin-Resistant Enterococcus | en_US |
dc.title | Development of a new multiplex real-time PCR assay for rapid screening of hospital-acquired infection agents | en_US |
dc.type | article | en_US |
dc.relation.ispartof | Journal of Microbiological Methods | en_US |
dc.department | İstanbul Medipol Üniversitesi, Tıp Fakültesi, Temel Tıp Bilimleri Bölümü, Tıbbi Mikrobiyoloji Ana Bilim Dalı | en_US |
dc.department | İstanbul Medipol Üniversitesi, Tıp Fakültesi, Dahili Tıp Bilimleri Bölümü, Enfeksiyon Hastalıkları ve Klinik Mikrobiyoloji Ana Bilim Dalı | en_US |
dc.authorid | 0000-0003-3952-1914 | en_US |
dc.authorid | 0000-0001-8022-7325 | en_US |
dc.identifier.volume | 206 | en_US |
dc.relation.publicationcategory | Makale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanı | en_US |
dc.identifier.doi | 10.1016/j.mimet.2023.106690 | en_US |
dc.institutionauthor | İstanbullu Tosun, Ayşe | |
dc.institutionauthor | Yılmaz, Mesut | |
dc.identifier.wosquality | Q3 | en_US |
dc.identifier.wos | 000949983800001 | en_US |
dc.identifier.scopus | 2-s2.0-85148672009 | en_US |
dc.identifier.pmid | 36801238 | en_US |
dc.identifier.scopusquality | Q3 | en_US |