Development of a new multiplex real-time PCR assay for rapid screening of hospital-acquired infection agents

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dc.authorid0000-0003-3952-1914
dc.authorid0000-0001-8022-7325
dc.contributor.authorİstanbullu Tosun, Ayşe
dc.contributor.authorKolukırık, Mustafa
dc.contributor.authorYılmaz, Mesut
dc.contributor.authorNar Ötgün, Selin
dc.contributor.authorAygün, Gökhan
dc.contributor.authorKetre Kolukırık, Canan Zöhre
dc.contributor.authorZeybek, Ümit
dc.contributor.authorGirgin Özgümüş, Gözde
dc.contributor.authorTuran, Meral
dc.contributor.authorKuşkucu, Mert
dc.contributor.authorİnce, Orhan
dc.contributor.authorİnce, Bahar
dc.contributor.authorKılıç, Selçuk
dc.date.accessioned2023-03-03T09:50:56Z
dc.date.available2023-03-03T09:50:56Z
dc.date.issued2023
dc.departmentİstanbul Medipol Üniversitesi, Tıp Fakültesi, Temel Tıp Bilimleri Bölümü, Tıbbi Mikrobiyoloji Ana Bilim Dalı
dc.departmentİstanbul Medipol Üniversitesi, Tıp Fakültesi, Dahili Tıp Bilimleri Bölümü, Enfeksiyon Hastalıkları ve Klinik Mikrobiyoloji Ana Bilim Dalı
dc.description.abstractAims: 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.
dc.identifier.citationİstanbullu 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
dc.identifier.doi10.1016/j.mimet.2023.106690
dc.identifier.issn0167-7012
dc.identifier.issn1872-8359
dc.identifier.pmid36801238
dc.identifier.scopus2-s2.0-85148672009
dc.identifier.scopusqualityQ3
dc.identifier.urihttps://dx.doi.org/10.1016/j.mimet.2023.106690
dc.identifier.urihttps://hdl.handle.net/20.500.12511/10563
dc.identifier.volume206
dc.identifier.wos000949983800001en_US
dc.identifier.wosqualityQ3
dc.indekslendigikaynakWeb of Science
dc.indekslendigikaynakScopus
dc.indekslendigikaynakPubMed
dc.institutionauthorİstanbullu Tosun, Ayşe
dc.institutionauthorYılmaz, Mesut
dc.language.isoen
dc.publisherElsevier B.V.
dc.relation.ispartofJournal of Microbiological Methodsen_US
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanı
dc.rightsinfo:eu-repo/semantics/embargoedAccess
dc.subjectCarbapenem-Resistant Enterobacteriaceae
dc.subjectHospital-Acquired Infection
dc.subjectMethicillin-Resistant Staphylococcus Aureus
dc.subjectReal-Time PCR
dc.subjectVancomycin-Resistant Enterococcus
dc.titleDevelopment of a new multiplex real-time PCR assay for rapid screening of hospital-acquired infection agents
dc.typeArticle

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