About: Evaluation of TaqMan qPCR System Integrating Two Identically Labelled Hydrolysis Probes in Single Assay

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About: Evaluation of TaqMan qPCR System Integrating Two Identically Labelled Hydrolysis Probes in Single Assay Goto Sponge NotDistinct Permalink

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

Attributes	Values
type	Academic Article research paper schema:ScholarlyArticle
isDefinedBy	Covid-on-the-Web dataset
has title	Evaluation of TaqMan qPCR System Integrating Two Identically Labelled Hydrolysis Probes in Single Assay
Creator	Havlíčková, Martina Horníčková, Jitka Jiřincová, Helena Křivda, Vlastimil Nagy, Alexander Sedlák, Kamil Vitásková, Eliška
Source	PMC
abstract	Ongoing evolution of viral pathogens is a significant issue in diagnostic virology employing TaqMan qPCR/RT-qPCR. Specific concerns are related to false negativity due to probe binding failure. One option for compensating for such deficiency is to integrate a second identically labelled probe in the assay. However, how this alteration influences the reaction parameters has not been comprehensively demonstrated. In the present study, we evaluate a TaqMan protocol using two identically labelled hydrolysis probes (simple, LNA (locked-nucleic-acid)) and MGB (minor-groove-binder) modified probes and combinations thereof in a single assay. Our results based on a synthetic amplicon suggest that the second probe does not compromise the TaqMan qPCR/RT-qPCR parameters, which repeatedly and reproducibly remained comparable to those of the corresponding single-probe assays, irrespective of the relative probe orientation, whether opposite or tandem, and probe modifications or combinations thereof. On the other hand, the second probe additively contributed to the overall fluorescence signal. The utility of the dual-probe approach was demonstrated on practical examples by using field specimens. We hope that the present study might serve as a theoretical basis for the development or improvement of TaqMan qPCR/RT-qPCR assays for the detection of highly variable nucleic acid templates.
has issue date	2017-01-25(xsd:dateTime)
bibo:doi	10.1038/srep41392
bibo:pmid	28120891
has license	cc-by
sha1sum (hex)	a7da39018f8bac38c77086632a5ab80e8fe73516
schema:url	https://doi.org/10.1038/srep41392
resource representing a document's title	Evaluation of TaqMan qPCR System Integrating Two Identically Labelled Hydrolysis Probes in Single Assay
has PubMed Central identifier	PMC5264587
has PubMed identifier	28120891
schema:publication	Sci Rep
resource representing a document's body	covid:a7da39018f8bac38c77086632a5ab80e8fe73516#body_text
is schema:about of	named entity 'assays' named entity 'based' named entity 'probe' named entity 'probes' named entity 'However' named entity 'RT-qPCR' named entity 'comparable' named entity 'EVALUATION' named entity 'PROBES' named entity 'LNA' named entity 'FIELD' named entity 'INTEGRATING' named entity 'FLUORESCENCE' named entity 'ORIENTATION' named entity 'REACTION' named entity 'OUR' named entity 'ASSAY' named entity 'TO INTEGRATE' named entity 'EVOLUTION' named entity 'DETECTION' named entity 'SINGLE' named entity 'VARIABLE' named entity 'HAND' named entity 'HOPE' named entity 'FALSE' named entity 'QPCR' named entity 'CONCERNS' named entity 'EVALUATE' named entity 'ALTERATION' named entity 'COMBINATIONS' named entity 'RELATIVE' named entity 'SUGGEST' named entity 'DEMONSTRATED' named entity 'AMPLICON' named entity 'COMPENSATING' named entity 'NEGATIVITY' named entity 'PROTOCOL' named entity 'GROOVE' named entity 'TAQMAN' named entity 'OVERALL' named entity 'PARAMETERS' named entity 'DEFICIENCY' named entity 'SECOND' named entity 'BASED' named entity 'RESULTS' named entity 'SYSTEM' named entity 'PROBES' named entity 'HYDROLYSIS' named entity 'OPEN' named entity 'TEMPLATES' named entity 'DIAGNOSTIC' named entity 'UTILITY' named entity 'HIGHLY' named entity 'TANDEM' named entity 'APPROACH' named entity 'BASIS' named entity 'REPEATEDLY' named entity 'SPECIFIC' named entity 'SYNTHETIC' named entity 'BINDER' named entity 'VIROLOGY' named entity 'NUCLEIC ACID' named entity 'USING' named entity 'IMPROVEMENT' named entity 'COMPROMISE' named entity 'SIGNIFICANT' named entity 'RELATED' named entity 'OPPOSITE' named entity 'SPECIMENS' named entity 'MODIFIED' named entity 'ONGOING' named entity 'BINDING' named entity 'VIRAL PATHOGENS' named entity 'OPTION' named entity 'SIGNAL'

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