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Naslov:Assessment of the real-time PCR and different digital PCR platforms for DNA quantification
Avtorji:ID Pavšič, Jernej (Avtor)
ID Žel, Jana (Avtor)
ID Milavec, Mojca (Avtor)
Datoteke:.pdf PDF - Predstavitvena datoteka, prenos (1,19 MB)
MD5: 2DF81B25A6A4AE63EB45FFFF6D6E404A
 
URL URL - Izvorni URL, za dostop obiščite https://doi.org/10.1007/s00216-015-9107-2
 
Jezik:Angleški jezik
Tipologija:1.01 - Izvirni znanstveni članek
Organizacija:Logo NIB - Nacionalni inštitut za biologijo
Povzetek:Digital PCR (dPCR) is beginning to supersede real-time PCR (qPCR) for quantification of nucleic acids in many different applications. Several analytical properties of the two most commonly used dPCR platforms, namely the QX100 system (Bio-Rad) and the 12.765 array of the Biomark system (Fluidigm), have already been evaluated and compared with those of qPCR. However, to the best of our knowledge, direct comparison between the three of these platforms using the same DNA material has not been done, and the 37 K array on the Biomark system has also not been evaluated in terms of linearity, analytical sensitivity and limit of quantification. Here, a first assessment of qPCR, the QX100 system and both arrays of the Biomark system was performed with plasmid and genomic DNA from human cytomegalovirus. With use of PCR components that alter the efficiency of qPCR, each dPCR platform demonstrated consistent copy-number estimations, which indicates the high resilience of dPCR. Two approaches, one considering the total reaction volume and the other considering the effective reaction size, were used to assess linearity, analytical sensitivity and variability. When the total reaction volume was considered, the best performance was observed with qPCR, followed by the QX100 system and the Biomark system. In contrast, when the effective reaction size was considered, all three platforms showed almost equal limits of detection and variability. Although dPCR might not always be more appropriate than qPCR for quantification of low copy numbers, dPCR is a suitable method for robust and reproducible quantification of viral DNA, and a promising technology for the higher-order reference measurement method.
Ključne besede:real-time PCR, molecular diagnostics, human cytomegalovirus, DNA quantification, digital PCR
Status publikacije:Objavljeno
Verzija publikacije:Objavljena publikacija
Datum objave:01.01.2016
Leto izida:2016
Št. strani:str. 107-121
Številčenje:Vol. 408, iss. 1
PID:20.500.12556/DiRROS-19699 Novo okno
UDK:577.2
ISSN pri članku:1618-2642
DOI:10.1007/s00216-015-9107-2 Novo okno
COBISS.SI-ID:3649615 Novo okno
Datum objave v DiRROS:25.07.2024
Število ogledov:365
Število prenosov:215
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:Drugi - Drug financer ali več financerjev
Program financ.:EURAMET and the European Union
Akronim:Infect-Met

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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