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About:
Detection and differentiation of field and vaccine strains of canine distemper virus using reverse transcription followed by nested real time PCR (RT-nqPCR) and RFLP analysis
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An Entity of Type :
schema:ScholarlyArticle
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covidontheweb.inria.fr
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Type:
Academic Article
research paper
schema:ScholarlyArticle
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type
Academic Article
research paper
schema:ScholarlyArticle
isDefinedBy
Covid-on-the-Web dataset
has title
Detection and differentiation of field and vaccine strains of canine distemper virus using reverse transcription followed by nested real time PCR (RT-nqPCR) and RFLP analysis
Creator
Ikuta, Nilo
Wageck Canal, Cláudio
Makiejczuk, Aline
Cardoso, Cristine
Da Costa Allgayer, Mariangela
Dossin, Cristine
Fischer, Bastos
Fonseca, Kazantzi
Lehmann, Fernanda
Lunge, Ricardo
Salvador, André
Source
Elsevier; Medline; PMC
abstract
Abstract Canine distemper virus (CDV) is the cause of a severe and highly contagious disease in dogs. Practical diagnosis of canine distemper based on clinical signs and laboratory tests are required to confirm CDV infection. The present study aimed to develop a molecular assay to detect and differentiate field and vaccine CDV strains. Reverse transcription followed by nested real time polymerase chain reaction (RT-nqPCR) was developed, which exhibited analytical specificity (all the samples from healthy dogs and other canine infectious agents were not incorrectly detected) and sensitivity (all replicates of a vaccine strain were positive up to the 3125-fold dilution – 100.7 TCID50). RT-nqPCR was validated for CDV detection on different clinical samples (blood, urine, rectal and conjunctival swabs) of 103 animals suspected to have distemper. A total of 53 animals were found to be positive based on RT-nqPCR in at least one clinical sample. Blood resulted in more positive samples (50 out of 53, 94.3%), followed by urine (44/53, 83.0%), rectal (38/53, 71%) and conjunctival (27/53, 50.9%) swabs. A commercial immunochromatography (IC) assay had detected CDV in only 30 conjunctival samples of these positive dogs. Nucleoprotein (NC) gene sequencing of 25 samples demonstrated that 23 of them were closer to other Brazilian field strains and the remaining two to vaccine strains. A single nucleotide sequences difference, which creates an Msp I restriction enzyme digestion, was used to differentiate between field and vaccine CDV strains by restriction fragment length polymorphism (RFLP) analysis. The complete assay was more sensitive than was IC for the detection of CDV. Blood was the more frequently positive specimen and the addition of a restriction enzyme step allowed the differentiation of vaccine and Brazilian field strains.
has issue date
2013-12-31
(
xsd:dateTime
)
bibo:doi
10.1016/j.jviromet.2013.08.002
bibo:pmid
23942341
has license
els-covid
sha1sum (hex)
2239c1dd1a7f18280cc59b62f43be30d25dd26a8
schema:url
https://doi.org/10.1016/j.jviromet.2013.08.002
resource representing a document's title
Detection and differentiation of field and vaccine strains of canine distemper virus using reverse transcription followed by nested real time PCR (RT-nqPCR) and RFLP analysis
has PubMed Central identifier
PMC7113657
has PubMed identifier
23942341
schema:publication
Journal of Virological Methods
resource representing a document's body
covid:2239c1dd1a7f18280cc59b62f43be30d25dd26a8#body_text
is
schema:about
of
named entity 'creates'
named entity 'highly'
named entity 'frequently'
named entity 'single'
named entity 'replicates'
named entity 'sequences'
named entity 'conjunctival'
named entity 'assay'
named entity 'CLINICAL SIGNS '
named entity 'REPLICATES'
named entity 'STEP'
named entity 'VALIDATED'
named entity 'SAMPLES'
named entity 'ALLOWED'
named entity 'CANINE DISTEMPER VIRUS'
named entity '0.7'
named entity 'GENE SEQUENCING'
named entity 'HIGHLY CONTAGIOUS DISEASE'
named entity 'SENSITIVE'
named entity 'USED'
named entity 'FOLD DILUTION'
named entity 'DIFFERENTIATE'
named entity 'MOLECULAR ASSAY'
named entity 'COMPLETE'
named entity 'DEMONSTRATED'
named entity 'ANIMALS'
named entity 'CDV'
named entity 'REVERSE TRANSCRIPTION'
named entity 'FIELD'
named entity 'DISTEMPER'
named entity 'BLOOD'
named entity 'STUDY'
named entity 'THESE'
named entity 'CLINICAL'
named entity 'ADDITION'
named entity 'HAVE'
named entity 'RESTRICTION ENZYME DIGESTION'
named entity 'SUSPECTED'
named entity 'TCID'
named entity 'BRAZILIAN'
named entity 'REMAINING'
named entity 'RESTRICTION ENZYME'
named entity 'NUCLEOTIDE SEQUENCES'
named entity 'CANINE DISTEMPER'
named entity 'DIFFERENT'
named entity 'HEALTHY'
named entity 'PRESENT'
named entity 'CANINE'
named entity 'UP TO'
named entity 'STRAINS'
named entity 'VACCINE'
named entity 'TOTAL'
named entity 'DIFFERENTIATION'
named entity 'MSP'
named entity 'DIAGNOSIS'
named entity 'NUCLEOPROTEIN'
named entity 'REQUIRED'
named entity 'DIFFERENCE'
named entity 'INFECTION'
named entity 'FREQUENTLY'
named entity 'SAMPLE'
named entity 'LABORATORY TESTS'
named entity 'COMMERCIAL'
named entity 'SPECIMEN'
named entity 'FOLLOWED BY'
named entity 'ANALYSIS'
named entity 'RECTAL'
named entity 'VACCINE STRAIN'
named entity 'DETECTED'
named entity 'SWABS'
named entity 'URINE'
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