About: Hot start reverse transcriptase: an approach for improved real-time RT-PCR performance

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An Entity of Type : schema:ScholarlyArticle, within Data Space : covidontheweb.inria.fr associated with source document(s)

Attributes	Values
type	Academic Article research paper schema:ScholarlyArticle
isDefinedBy	Covid-on-the-Web dataset
title	Hot start reverse transcriptase: an approach for improved real-time RT-PCR performance
Creator	Klöck, Gerd Leune, Annika Möller, Ronny Rutschke, Nils Winterhalter, Mathias Zimmermann, Jan
source	Medline; PMC
abstract	BACKGROUND: Reverse transcriptase is an indispensable enzyme for real-time reverse transcriptase (RT)-PCR, a standard method in molecular diagnostics for detection and quantification of defined RNA molecules. The prevention of non-specific products due to elongation of misprimed oligonucleotides by the enzyme at temperatures beneath the specific annealing temperature is one of the biggest challenges in real-time RT-PCR. In the present study, an aptamer directed against the reverse transcriptase was analyzed for its potential to attain a temperature-dependent reverse transcriptase (“hot start” RT). FINDINGS: The hot start effect was investigated in a one-step real-time RT-PCR assay for the detection of Middle East respiratory syndrome coronavirus (MERS-CoV). Results with aptamer revealed a reduced RT activity at low temperatures while achieving full activity at the specific annealing temperature of 55 °C. Sensitivity (limit of detection (LoD) 95 %) of the MERS-CoV assay was increased by about two times in the presence of aptamer. CONCLUSIONS: The study demonstrates the potential of aptamer-dependent hot start RT for the improvement of diagnostic real-time RT-PCR assays.
has issue date	2015-06-21(xsd:dateTime)
bibo:doi	10.1186/s40543-015-0063-4
bibo:pmid	32226638
has license	cc-by
sha1sum (hex)	f9c159585ae628a508b58c57fd9b23d4973d4b95
schema:url	https://doi.org/10.1186/s40543-015-0063-4
resource representing a document's title	Hot start reverse transcriptase: an approach for improved real-time RT-PCR performance
has PubMed Central identifier	PMC7099353
has PubMed identifier	32226638
schema:publication	J Anal Sci Technol
resource representing a document's body	covid:f9c159585ae628a508b58c57fd9b23d4973d4b95#body_text
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