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Naslov:Development of novel digital PCR assays for the rapid quantification of Gram-negative bacteria biomarkers using RUCS algorithm
Avtorji:ID Bogožalec Košir, Alexandra (Avtor)
ID Alič, Špela (Avtor)
ID Tomič, Viktorija (Avtor)
ID Lužnik, Dane (Avtor)
ID Dreo, Tanja (Avtor)
ID Milavec, Mojca (Avtor)
Datoteke:URL URL - Izvorni URL, za dostop obiščite https://doi.org/10.1016/j.ymeth.2024.10.011
 
.pdf PDF - Predstavitvena datoteka, prenos (2,31 MB)
MD5: 91FD8FB54941BE1D49655A318AED9464
 
Jezik:Angleški jezik
Tipologija:1.01 - Izvirni znanstveni članek
Organizacija:Logo NIB - Nacionalni inštitut za biologijo
Povzetek:Rapid and accurate identification of bacterial pathogens is crucial for effective treatment and infection control, particularly in hospital settings. Conventional methods like culture techniques and MALDI-TOF mass spectrometry are often time-consuming and less sensitive. This study addresses the need for faster and more precise diagnostic methods by developing novel digital PCR (dPCR) assays for the rapid quantification of biomarkers from three Gram-negative bacteria: Acinetobacter baumannii, Klebsiella pneumoniae, and Pseudomonas aeruginosa. Utilizing publicly available genomes and the rapid identification of PCR primers for unique core sequences or RUCS algorithm, we designed highly specific dPCR assays. These assays were validated using synthetic DNA, bacterial genomic DNA, and DNA extracted from clinical samples. The developed dPCR methods demonstrated wide linearity, a low limit of detection (approx. 30 copies per reaction), and robust analytical performance with measurement uncertainty below 25 %. The assays showed high repeatability and intermediate precision, with no cross-reactivity observed. Comparison with MALDI-TOF mass spectrometry revealed substantial concordance, highlighting the methods’ suitability for clinical diagnostics. This study underscores the potential of dPCR for rapid and precise quantification of Gram-negative bacterial biomarkers. The developed methods offer significant improvements over existing techniques, providing faster, more accurate, and SI-traceable measurements. These advancements could enhance clinical diagnostics and infection control practices.
Ključne besede:digital PCR (dPCR), Gram-negative bacteria, pathogen detection, respiratory infections, biomarkers, RUCS algorithm
Status publikacije:Objavljeno
Verzija publikacije:Objavljena publikacija
Datum objave:01.12.2024
Leto izida:2024
Št. strani:str. 72-80
Številčenje:Vol. 232
PID:20.500.12556/DiRROS-20758 Novo okno
UDK:579
ISSN pri članku:1046-2023
DOI:10.1016/j.ymeth.2024.10.011 Novo okno
COBISS.SI-ID:213682691 Novo okno
Opomba:Soavtorji: Špela Alič, Viktorija Tomič, Dane Lužnik, Tanja Dreo, Mojca Milavec;
Datum objave v DiRROS:05.11.2024
Število ogledov:312
Število prenosov:431
Metapodatki:XML DC-XML DC-RDF
:
BOGOŽALEC KOŠIR, Alexandra, ALIČ, Špela, TOMIČ, Viktorija, LUŽNIK, Dane, DREO, Tanja in MILAVEC, Mojca, 2024, Development of novel digital PCR assays for the rapid quantification of Gram-negative bacteria biomarkers using RUCS algorithm. Methods [na spletu]. 2024. Vol. 232, p. 72–80. [Dostopano 9 april 2025]. DOI 10.1016/j.ymeth.2024.10.011. Pridobljeno s: https://dirros.openscience.si/IzpisGradiva.php?lang=slv&id=20758
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Gradivo je del revije

Naslov:Methods
Skrajšan naslov:Methods
Založnik:Academic Press
ISSN:1046-2023
COBISS.SI-ID:862740 Novo okno

Gradivo je financirano iz projekta

Financer:ARIS - Javna agencija za znanstvenoraziskovalno in inovacijsko dejavnost Republike Slovenije
Številka projekta:Z2-1860-2019
Naslov:Meroslovje za sledenje bolnišničnih okužb respiratornega trakta, ki jih povzročajo Gram-negativne bakterije
Financer:ARIS - Javna agencija za znanstvenoraziskovalno in inovacijsko dejavnost Republike Slovenije
Številka projekta:P4-0165-2022
Naslov:Biotehnologija in sistemska biologija rastlin
Financer:EC - European Commission
Program financ.:SEPTIMET
Financer:Drugi - Drug financer ali več financerjev
Program financ.:European Union, European Regional Development Fund

Licence

Licenca:CC BY 4.0, Creative Commons Priznanje avtorstva 4.0 Mednarodna
Povezava:http://creativecommons.org/licenses/by/4.0/deed.sl
Opis:To je standardna licenca Creative Commons, ki daje uporabnikom največ možnosti za nadaljnjo uporabo dela, pri čemer morajo navesti avtorja.

Sekundarni jezik

Jezik:Slovenski jezik
Ključne besede:digitalna PCR, Gram-negativne bakterije, detekcija patogenov, okužba dihal, biomarkerji, diagnostika, molekularna biologija


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