Quantitative one-step real-time RT-PCR for the fast detection of the four genotypes of PPRV. 2010

Olivier Kwiatek, and Djénéba Keita, and Patricia Gil, and Jovita Fernández-Pinero, and Miguel Angel Jimenez Clavero, and Emmanuel Albina, and Genevieve Libeau
CIRAD, UMR CIRAD/INRA Contrôle des Maladies, F-34398 Montpellier Cedex 5, France.

A one-step real-time Taqman RT-PCR assay (RRT-PCR) for peste des petits ruminants virus (PPRV) was developed to detect the four lineages of PPRV by targeting the nucleoprotein (N) gene of the virus. This new assay was compared to a conventional RT-PCR on reference strains and field materials. Quantitation was performed against a standard based on a synthetic transcript of the NPPR gene for which a minimum of 32 copies per reaction were detected with a corresponding C(t) value of 39. Depending on the lineage involved, the detection limit of RRT-PCR was decreased by one to three log copies relative to the conventional method. The lower stringency occurred with lineage III because of minor nucleotide mismatches within the probe region. The assay did not detect phylogenetically or symptomatically related viruses of ruminants (such as rinderpest, bluetongue, and bovine viral diarrhea viruses). However, it was capable of detecting 20% more positive field samples with low viral RNA loads compared to the conventional PCR method. When compared on a proficiency panel to the method developed by Bao et al. (2008), the sensitivity of the in-house assay was slightly improved on lineage II. It proved significantly faster to perform and hence better adapted for monitoring large numbers of at risk or diseased animals.

UI MeSH Term Description Entries
D002162 Camelus Two-toed, hoofed mammals with four legs, a big-lipped snout, and a humped back belonging to the family Camelidae. They are native to North Africa, and Western and Central Asia. Camels,Dromedary,Bactrian Camels,Bractrian Camels,Camelus bactrianus,Camelus dromedarius,Bactrian Camel,Bractrian Camel,Camel,Camel, Bactrian,Camel, Bractrian,Camels, Bactrian,Camels, Bractrian,Dromedaries
D003937 Diagnosis, Differential Determination of which one of two or more diseases or conditions a patient is suffering from by systematically comparing and contrasting results of diagnostic measures. Diagnoses, Differential,Differential Diagnoses,Differential Diagnosis
D006041 Goats Any of numerous agile, hollow-horned RUMINANTS of the genus Capra, in the family Bovidae, closely related to the SHEEP. Capra,Capras,Goat
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D012680 Sensitivity and Specificity Binary classification measures to assess test results. Sensitivity or recall rate is the proportion of true positives. Specificity is the probability of correctly determining the absence of a condition. (From Last, Dictionary of Epidemiology, 2d ed) Specificity,Sensitivity,Specificity and Sensitivity
D012756 Sheep Any of the ruminant mammals with curved horns in the genus Ovis, family Bovidae. They possess lachrymal grooves and interdigital glands, which are absent in GOATS. Ovis,Sheep, Dall,Dall Sheep,Ovis dalli
D012757 Sheep Diseases Diseases of domestic and mountain sheep of the genus Ovis. Ovine Diseases,Disease, Ovine,Disease, Sheep,Diseases, Ovine,Diseases, Sheep,Ovine Disease,Sheep Disease
D015511 Goat Diseases Diseases of the domestic or wild goat of the genus Capra. Caprine Diseases,Caprine Disease,Disease, Caprine,Disease, Goat,Diseases, Caprine,Diseases, Goat,Goat Disease
D018107 Peste-des-petits-ruminants virus A species of MORBILLIVIRUS causing a severe, often fatal enteritis and pneumonia (PESTE-DES-PETITS-RUMINANTS) in sheep and goats. Peste des petits ruminants virus
D019562 Viral Load The quantity of measurable virus in a body fluid. Change in viral load, measured in plasma, is sometimes used as a SURROGATE MARKER in disease progression. Viral Burden,Virus Titer,Burden, Viral,Load, Viral,Titer, Virus

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