About: Development of a one-step real-time quantitative PCR assay based on primer-probe energy transfer for the detection of porcine reproductive and respiratory syndrome virus

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About: Development of a one-step real-time quantitative PCR assay based on primer-probe energy transfer for the detection of porcine reproductive and respiratory syndrome virus Goto Sponge NotDistinct Permalink

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

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type	Academic Article research paper schema:ScholarlyArticle
isDefinedBy	Covid-on-the-Web dataset
has title	Development of a one-step real-time quantitative PCR assay based on primer-probe energy transfer for the detection of porcine reproductive and respiratory syndrome virus
Creator	Benyeda, Zsófia Balka, Gyula Bálint, Ádám Hornyák, Ákos Rusvai, Miklós
Source	Elsevier; Medline; PMC
abstract	Abstract A one-step real-time RT-PCR method has been developed for the simultaneous detection of both genotypes of porcine reproductive and respiratory syndrome virus (PRRSV). The assay is based on primer-probe energy transfer, and the most important advantage of this is the relative tolerance towards mutations in the target-probe region. The primers and the probe were designed using an alignment of 235 Type 1 (including all subtypes) and Type 2 PRRSV strains. According to the alignment, multiple degenerations were included in the forward and reverse primers to enable the detection of all PRRSV strains deposited in the GenBank. Specificity was tested using 37 different PRRSV strains and eight other swine pathogen viruses. The detection limit was approximately 10 copies of RNA prepared from the Lelystad virus, a European Subtype 3 virus (Belarus strain Soz-8), and an American vaccine virus (Ingelvac MLV®). One TCID50 was the detection limit in the case of the cell cultured Lelystad virus and an American wild type isolate, respectively. The melting point analysis revealed melting point decrease, but no significant sensitivity and signal loss in the presence of numerous (up to five) target-probe mismatches, indicating the capability of tolerating even more mutations. The method was suitable for the detection and quantitation of phylogenetically divergent strains and can serve as a robust, high throughput tool for molecular diagnosis of the PRRSV.
has issue date	2009-06-30(xsd:dateTime)
bibo:doi	10.1016/j.jviromet.2009.01.014
bibo:pmid	19200443
has license	els-covid
sha1sum (hex)	d8a9831c00106017c617e0491df9dba67c19f115
schema:url	https://doi.org/10.1016/j.jviromet.2009.01.014
resource representing a document's title	Development of a one-step real-time quantitative PCR assay based on primer-probe energy transfer for the detection of porcine reproductive and respiratory syndrome virus
has PubMed Central identifier	PMC7112897
has PubMed identifier	19200443
schema:publication	Journal of Virological Methods
resource representing a document's body	covid:d8a9831c00106017c617e0491df9dba67c19f115#body_text
is schema:about of	named entity 'REAL-TIME QUANTITATIVE PCR' named entity 'deposited' named entity 'enable' named entity 'virus' named entity 'strains' named entity 'European' named entity 'reverse' named entity 'reproductive' named entity 'DEVELOPMENT' named entity 'ADVANTAGE' named entity 'SIGNIFICANT' named entity 'DEGENERATIONS' named entity 'TARGET' named entity 'REVERSE' named entity 'ENERGY TRANSFER' named entity 'SPECIFICITY' named entity 'DECREASE' named entity 'BUT' named entity 'PREPARED' named entity 'ANALYSIS' named entity 'RELATIVE TOLERANCE' named entity 'PORCINE REPRODUCTIVE AND RESPIRATORY SYNDROME VIRUS' named entity 'HIGH THROUGHPUT' named entity 'TCID' named entity 'NUMEROUS' named entity 'MLV' named entity 'APPROXIMATELY' named entity 'CAPABILITY' named entity 'PROBE' named entity 'SUBTYPES' named entity 'TYPE 2' named entity 'PORCINE REPRODUCTIVE AND RESPIRATORY SYNDROME VIRUS' named entity 'CULTURED' named entity 'REAL-TIME RT-PCR' named entity 'STRAIN' named entity 'VIRUS' named entity 'MULTIPLE' named entity 'TESTED' named entity 'ISOLATE' named entity 'DETECTION' named entity 'QUANTITATION' named entity 'REVEALED' named entity 'DEPOSITED' named entity 'FORWARD' named entity 'BASED' named entity 'ENERGY TRANSFER' named entity 'INCLUDED' named entity 'PRESENCE OF' named entity 'RNA' named entity 'CASE' named entity 'STRAINS' named entity 'SENSITIVITY' named entity 'ENABLE' named entity 'SWINE' named entity 'IMPORTANT' named entity 'ACCORDING' named entity 'ASSAY' covid:arg/d8a9831c00106017c617e0491df9dba67c19f115 named entity 'WILD TYPE' named entity 'PATHOGEN' named entity 'MOLECULAR DIAGNOSIS' named entity 'PRIMERS' named entity 'VACCINE' named entity 'MUTATIONS' named entity 'USING' named entity 'PRIMER' named entity 'BASED' named entity 'TOOL' named entity 'COPIES' named entity 'AMERICAN' named entity 'SIGNAL LOSS' named entity 'SUITABLE' named entity 'REGION' named entity 'EUROPEAN SUBTYPE' named entity 'INDICATING' named entity 'MELTING POINT' named entity 'GENOTYPES'

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