Validation of two commercial real-time RT-PCR kits for rapid and specific diagnosis of classical swine fever virus

• Published in: Journal of virological methods Vol. 147; no. 1; pp. 136 - 142
• Main Authors: Le Dimna, M., Vrancken, R., Koenen, F., Bougeard, S., Mesplède, A., Hutet, E., Kuntz-Simon, G., Le Potier, M.F.
• Format: Journal Article
• Language: English
• Published: London Elsevier B.V 2008; Amsterdam Elsevier; New York, NY
• Subjects: Animals; Biological and medical sciences; Classical swine fever; Classical Swine Fever - diagnosis; Classical Swine Fever - virology; Classical swine fever virus; Classical swine fever virus - genetics; Classical swine fever virus - isolation & purification; Diagnosis validation; Fundamental and applied biological sciences. Psychology; Microbiology; Pestivirus; Real-time RT-PCR; Reproducibility of Results; Reverse Transcriptase Polymerase Chain Reaction - methods; Sensitivity and Specificity; Swine; Techniques used in virology; Virology; Real-time RT-PCR; Diagnosis validation; Classical swine fever; Pestivirus; Microbiology; Method; Real time; Virology; Infection; Virus; Viral disease; Swine fever; Diagnosis; Flaviviridae; Veterinary; Reverse transcription polymerase chain reaction; Hog cholera virus
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/j.jviromet.2007.08.013

Two real-time RT-PCR kits, developed by LSI (TaqVet CSF) and ADIAGENE (Adiavet CSF), obtained an agreement to be commercialised in France, subject to conditions, defined by the French Classical Swine Fever (CSF) National Reference Laboratory. The producers were asked to introduce an internal control to check the RNA extraction efficacy. The different criteria assessed were sensitivity, “pestivirus specificity”, reproducibility and ease of handling, using 189 different samples. These samples were either CSFV inactivated strains or blood/serum/organs collected from CSFV experimentally infected pigs or naturally infected wild boars. The reproducibility of the assays was confirmed by the analysis of a batch-to-batch panel control that was used for inter-laboratory tests involving nine laboratories. The two kits were also tested for the use in mass diagnostics and the results proved the kits to be suited using pools of blood, serum and tonsils. Moreover, a field evaluation, carried out on spleen samples collected from the CSF surveillance of wild boars in an area known to be infected and from domestic pigs at a slaughterhouse, confirmed the high sensitivity and specificity of the two kits. This step-by-step evaluation procedure confirmed that the two commercial CSF real-time RT-PCR kits have a higher predictive value than the current diagnostic standard, Virus Isolation.