Evaluation of six TaqMan RT-rtPCR kits on two thermocyclers for the reliable detection of rabies virus RNA
| Autor: | Florence Cliquet, Evelyne Picard-Meyer, Carine Peytavin de Garam, J.L. Schereffer, Emmanuelle Robardet |
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| Přispěvatelé: | Laboratoire de la rage et de la faune sauvage de Nancy (LRFSN), Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES) |
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
TaqMan RT-rtPCR
Rabies 040301 veterinary sciences [SDV]Life Sciences [q-bio] Global Health medicine.disease_cause Sensitivity and Specificity 0403 veterinary science 03 medical and health sciences medicine TaqMan Animals Rabies virus RNA Full Scientific Reports Direct fluorescent antibody Lyssavirus 0303 health sciences [SDV.BA.MVSA]Life Sciences [q-bio]/Animal biology/Veterinary medicine and animal Health General Veterinary biology Reverse Transcriptase Polymerase Chain Reaction 030306 microbiology Rabies virus 04 agricultural and veterinary sciences medicine.disease biology.organism_classification Virology 3. Good health Real-time polymerase chain reaction [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology RNA Viral Immunohistochemistry Reagent Kits Diagnostic |
| Zdroj: | Journal of Veterinary Diagnostic Investigation Journal of Veterinary Diagnostic Investigation, American Association of Veterinary Laboratory Diagnosticians, 2019, 31 (1), pp.47-57. ⟨10.1177/1040638718818223⟩ |
| ISSN: | 1040-6387 |
| Popis: | Rabies is diagnosed postmortem in animals, based on tests prescribed by the World Organization for Animal Health (OIE), such as the fluorescent antibody test, the direct rapid immunohistochemistry test, or pan-lyssavirus PCR assays. Several reverse-transcription real-time PCR (RT-rtPCR) methods have been developed and validated for rapid and accurate detection of lyssaviruses. We evaluated the performance of 6 TaqMan RT-rtPCR kits using different commercial master mixes and 2 real-time thermocyclers. Changing the master mix overall did not influence the TaqMan RT-rtPCR performance, regardless of the thermocycler used. The limits of detection at the 95% confidence level were 18.1–25.8 copies/µL for the Rotor-Gene Q MDx thermocycler and 16.7–21.5 for the Mx3005P thermocycler. Excellent repeatability was demonstrated for rabies virus (RABV) RNA samples of 100, 50, and 25 copies/µL regardless of the thermocycler used. RABV field samples ( n = 35) isolated worldwide gave positive results using the most efficient of the 6 kits tested, with a copy number of 6.03 × 102 to 6.78 × 107 RNA copies per reaction. The TaqMan RT-rtPCR assay provides sensitive and rapid amplification of RABV RNA. |
| Databáze: | OpenAIRE |
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