Rapid Detection of Peste des Petits Ruminants Virus (PPRV) Nucleic Acid Using a Novel Low-Cost Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) Assay for Future Use in Nascent PPR Eradication Programme
| Autor: | John Flannery, Emma L.A. Howson, Mana Mahapatra, Muneeswaran Selvaraj, Carrie Batten, Veronica L. Fowler, Satya Parida |
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| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
040301 veterinary sciences lcsh:QR1-502 Loop-mediated isothermal amplification Sheep Diseases Biology Nose Eye Rapid detection Sensitivity and Specificity lcsh:Microbiology Article Disease Outbreaks Peste-des-petits-ruminants virus 0403 veterinary science 03 medical and health sciences Virology eradication programme Peste-des-Petits-Ruminants diagnostics Animals Pathology Molecular Reverse Transcription Loop-mediated Isothermal Amplification RT-LAMP DNA Primers Goat Diseases Sheep Goats Temperature 04 agricultural and veterinary sciences Reverse Transcription Nucleocapsid Proteins 3. Good health Vaccination 030104 developmental biology Infectious Diseases rapid detection DNA Viral Nucleic acid RNA extraction PPRV Nucleic Acid Amplification Techniques pen side |
| Zdroj: | Viruses Volume 11 Issue 8 Viruses, Vol 11, Iss 8, p 699 (2019) |
| ISSN: | 1999-4915 |
| Popis: | Peste des petits ruminants (PPR) is a disease of small ruminants caused by peste des petits ruminants virus (PPRV), and is endemic in Asia, the Middle East and Africa. Effective control combines the application of early warning systems, accurate laboratory diagnosis and reporting, animal movement restrictions, suitable vaccination and surveillance programs, and the coordination of all these measures by efficient veterinary services. Molecular assays, including conventional reverse transcription-polymerase chain reaction (RT-PCR) and real-time RT-PCR (RT-qPCR) have improved the sensitivity and rapidity of diagnosing PPR. However, currently these assays are only performed within laboratory settings therefore, the development of field diagnostics for PPR would improve the fast implementation of control policies, particularly when PPR has been targeted to be eradicated by 2030. Loop-mediated isothermal amplification (LAMP) assays are simple to use, rapid, and have sensitivity and specificity within the range of RT-qPCR and can be performed in the field using disposable consumables and portable equipment. This study describes the development of a novel RT-LAMP assay for the detection of PPRV nucleic acid by targeting the N-protein gene. The RT-LAMP assay was evaluated using cell culture propagated PPRVs, field samples from clinically infected animals and samples from experimentally infected animals encompassing all four lineages (I-IV) of PPRV. The test displayed 100% concordance with RT-qPCR when considering an RT-qPCR cut-off value of CT > 40. Further, the RT-LAMP assay was evaluated using experimental and outbreak samples without prior RNA extraction making it more time and cost-effective. This assay provides a solution for a pen-side, rapid and inexpensive PPR diagnostic for use in the field in nascent PPR eradication programme. |
| Databáze: | OpenAIRE |
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