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Naslov:Inter-laboratory assessment of different digital PCR platforms for quantification of human cytomegalovirus DNA
Avtorji:ID Pavšič, Jernej (Avtor)
ID Devonshire, Alison S. (Avtor)
ID Blejec, Andrej (Avtor)
ID Foy, Carole A. (Avtor)
ID Heuverswyn, Fran Van (Avtor)
ID Jones, Gerwyn M. (Avtor)
ID Schimmel, Heinz (Avtor)
ID Žel, Jana (Avtor)
ID Huggett, Jim F. (Avtor)
ID Redshaw, Nicholas (Avtor)
ID Karczmarczyk, Maria (Avtor)
ID Mozioglu, Erkan (Avtor)
ID Akyürek, Sema (Avtor)
ID Akgöz, Müslüm (Avtor)
ID Milavec, Mojca (Avtor)
Datoteke:URL URL - Izvorni URL, za dostop obiščite http://dx.doi.org/10.1007/s00216-017-0206-0
 
.pdf PDF - Predstavitvena datoteka, prenos (638,49 KB)
MD5: 315D9D64042B8F34F5B431556FF38B60
 
Jezik:Angleški jezik
Tipologija:1.01 - Izvirni znanstveni članek
Organizacija:Logo NIB - Nacionalni inštitut za biologijo
Povzetek:Quantitative PCR (qPCR) is an important tool in pathogen detection. However, the use of different qPCR components, calibration materials and DNA extraction methods reduces comparability between laboratories, which can result in false diagnosis and discrepancies in patient care. The wider establishment of a metrological framework for nucleic acid tests could improve the degree of standardisation of pathogen detection and the quantification methods applied in the clinical context. To achieve this, accurate methods need to be developed and implemented as reference measurement procedures, and to facilitate characterisation of suitable certified reference materials. Digital PCR (dPCR) has already been used for pathogen quantification by analysing nucleic acids. Although dPCR has the potential to provide robust and accurate quantification of nucleic acids, further assessment of its actual performance characteristics is needed before it can be implemented in a metrological framework, and to allow adequate estimation of measurement uncertainties. Here, four laboratories demonstrated reproducibility (expanded measurement uncertainties below 15%) of dPCR for quantification of DNA from human cytomegalovirus, with no calibration to a common reference material. Using whole-virus material and extracted DNA, an intermediate precision (coefficients of variation below 25%) between three consecutive experiments was noted. Furthermore, discrepancies in estimated mean DNA copy number concentrations between laboratories were less than twofold, with DNA extraction as the main source of variability. These data demonstrate that dPCR offers a repeatable and reproducible method for quantification of viral DNA, and due to its satisfactory performance should be considered as candidate for reference methods for implementation in a metrological framework.
Ključne besede:digital PCR, DNA quantification, inter-laboratory assessment, human cytomegalovirus, virus reference materials
Status publikacije:Objavljeno
Verzija publikacije:Objavljena publikacija
Datum objave:26.01.2017
Leto izida:2017
Št. strani:str. 2601-2614
Številčenje:Vol. 409, iss. 10
PID:20.500.12556/DiRROS-19697 Novo okno
UDK:578
ISSN pri članku:1618-2642
DOI:10.1007/s00216-017-0206-0 Novo okno
COBISS.SI-ID:4223055 Novo okno
Datum objave v DiRROS:24.07.2024
Število ogledov:398
Število prenosov:233
Metapodatki:XML DC-XML DC-RDF
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Gradivo je del revije

Naslov:Analytical and bioanalytical chemistry
Skrajšan naslov:Anal. bioanal. chem.
Založnik:Springer
ISSN:1618-2642
COBISS.SI-ID:6966294 Novo okno

Gradivo je financirano iz projekta

Financer:ARIS - Javna agencija za znanstvenoraziskovalno in inovacijsko dejavnost Republike Slovenije
Številka projekta:P4-0165-2015
Naslov:Biotehnologija in sistemska biologija rastlin

Financer:EC - European Commission
Program financ.:EURAMET and the European Union
Naslov:EMRP project
Akronim:INFECT MET

Financer:ARIS - Javna agencija za znanstvenoraziskovalno in inovacijsko dejavnost Republike Slovenije
Številka projekta:1000-13-0105

Financer:Drugi - Drug financer ali več financerjev
Program financ.:Metrology Institute of the Republic of Slovenia, with financial support from the 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.

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