Development and Evaluation of Molecular Pen-Side Assays without Prior RNA Extraction for Peste des Petits Ruminants (PPR) and Foot and Mouth Disease (FMD)
Animal diseases such as peste des petits ruminants (PPR) and foot and mouth disease (FMD) cause significant economic losses in endemic countries and fast, accurate in-field diagnostics would assist with surveillance and outbreak control. The detection of these pathogens is usually performed at refer...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-04-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/14/4/835 |
| _version_ | 1797408990809817088 |
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| author | David Edge Mana Mahapatra Shona Strachan James Turton Ryan Waters Camilla Benfield Nathan Nazareth Felix Njeumi Nelson Nazareth Satya Parida |
| author_facet | David Edge Mana Mahapatra Shona Strachan James Turton Ryan Waters Camilla Benfield Nathan Nazareth Felix Njeumi Nelson Nazareth Satya Parida |
| author_sort | David Edge |
| collection | DOAJ |
| description | Animal diseases such as peste des petits ruminants (PPR) and foot and mouth disease (FMD) cause significant economic losses in endemic countries and fast, accurate in-field diagnostics would assist with surveillance and outbreak control. The detection of these pathogens is usually performed at reference laboratories, tested using assays that are recommended by The World Organisation for Animal Health (OIE), leading to delays in pathogen detection. This study seeks to demonstrate a proof-of-concept approach for a molecular diagnostic assay that is compatible with material direct from nasal swab sampling, without the need for a prior nucleic acid extraction step, that could potentially be applied at pen-side for both PPR and FMD. The use of such a rapid, low-cost assay without the need for a cold chain could permit testing capacity to be established in remote, resource limited areas and support the surveillance activities necessary to meet the goal of eradication of PPR by 2030. Two individual assays were developed that detect > 99% of PPR and FMD sequences available in GenBank, demonstrating pan-serotype FMD and pan-lineage PPR assays. The ability for the BioGene XF reagent that was used in this study to lyse FMD and PPR viruses and amplify their nucleic acids in the presence of unprocessed nasal swab eluate was evaluated. The reagent was shown to be capable of detecting the viral RNA present in nasal swabs collected from naïve and infected target animals. A study was performed comparing the relative specificity and sensitivity of the new assays to the reference assays. The study used nasal swabs collected from animals before and after infection (12 cattle infected with FMDV and 5 goats infected with PPRV) and both PPR and FMD viral RNA were successfully detected two to four days post-infection in all animals using either the XF or reference assay reagents. These data suggest that the assays are at least as sensitive as the reference assays and support the need for further studies in a field setting. |
| first_indexed | 2024-03-09T04:06:41Z |
| format | Article |
| id | doaj.art-86ba7abb9ffd45629b04ac4910bf04d3 |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-09T04:06:41Z |
| publishDate | 2022-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-86ba7abb9ffd45629b04ac4910bf04d32023-12-03T14:04:45ZengMDPI AGViruses1999-49152022-04-0114483510.3390/v14040835Development and Evaluation of Molecular Pen-Side Assays without Prior RNA Extraction for Peste des Petits Ruminants (PPR) and Foot and Mouth Disease (FMD)David Edge0Mana Mahapatra1Shona Strachan2James Turton3Ryan Waters4Camilla Benfield5Nathan Nazareth6Felix Njeumi7Nelson Nazareth8Satya Parida9BioGene Limited, 6 The Business Centre, Harvard Way, Kimbolton PE28 0NJ, UKThe Pirbright Institute, Ash Road, Pirbright, Woking, Surrey GU24 ONF, UKThe Pirbright Institute, Ash Road, Pirbright, Woking, Surrey GU24 ONF, UKBioGene Limited, 6 The Business Centre, Harvard Way, Kimbolton PE28 0NJ, UKThe Pirbright Institute, Ash Road, Pirbright, Woking, Surrey GU24 ONF, UKRoyal Veterinary College, University of London, Hawkshead Lane, North Mimms, Hatfield AL9 7TA, UKBioGene Limited, 6 The Business Centre, Harvard Way, Kimbolton PE28 0NJ, UKFood and Agriculture Organization of the United Nations (FAO), Viale delle Terme di Caracalla, 00153 Rome, ItalyBioGene Limited, 6 The Business Centre, Harvard Way, Kimbolton PE28 0NJ, UKThe Pirbright Institute, Ash Road, Pirbright, Woking, Surrey GU24 ONF, UKAnimal diseases such as peste des petits ruminants (PPR) and foot and mouth disease (FMD) cause significant economic losses in endemic countries and fast, accurate in-field diagnostics would assist with surveillance and outbreak control. The detection of these pathogens is usually performed at reference laboratories, tested using assays that are recommended by The World Organisation for Animal Health (OIE), leading to delays in pathogen detection. This study seeks to demonstrate a proof-of-concept approach for a molecular diagnostic assay that is compatible with material direct from nasal swab sampling, without the need for a prior nucleic acid extraction step, that could potentially be applied at pen-side for both PPR and FMD. The use of such a rapid, low-cost assay without the need for a cold chain could permit testing capacity to be established in remote, resource limited areas and support the surveillance activities necessary to meet the goal of eradication of PPR by 2030. Two individual assays were developed that detect > 99% of PPR and FMD sequences available in GenBank, demonstrating pan-serotype FMD and pan-lineage PPR assays. The ability for the BioGene XF reagent that was used in this study to lyse FMD and PPR viruses and amplify their nucleic acids in the presence of unprocessed nasal swab eluate was evaluated. The reagent was shown to be capable of detecting the viral RNA present in nasal swabs collected from naïve and infected target animals. A study was performed comparing the relative specificity and sensitivity of the new assays to the reference assays. The study used nasal swabs collected from animals before and after infection (12 cattle infected with FMDV and 5 goats infected with PPRV) and both PPR and FMD viral RNA were successfully detected two to four days post-infection in all animals using either the XF or reference assay reagents. These data suggest that the assays are at least as sensitive as the reference assays and support the need for further studies in a field setting.https://www.mdpi.com/1999-4915/14/4/835pen-side PCRpeste des petits ruminantsfoot and mouth diseasePPRFMDPCR |
| spellingShingle | David Edge Mana Mahapatra Shona Strachan James Turton Ryan Waters Camilla Benfield Nathan Nazareth Felix Njeumi Nelson Nazareth Satya Parida Development and Evaluation of Molecular Pen-Side Assays without Prior RNA Extraction for Peste des Petits Ruminants (PPR) and Foot and Mouth Disease (FMD) Viruses pen-side PCR peste des petits ruminants foot and mouth disease PPR FMD PCR |
| title | Development and Evaluation of Molecular Pen-Side Assays without Prior RNA Extraction for Peste des Petits Ruminants (PPR) and Foot and Mouth Disease (FMD) |
| title_full | Development and Evaluation of Molecular Pen-Side Assays without Prior RNA Extraction for Peste des Petits Ruminants (PPR) and Foot and Mouth Disease (FMD) |
| title_fullStr | Development and Evaluation of Molecular Pen-Side Assays without Prior RNA Extraction for Peste des Petits Ruminants (PPR) and Foot and Mouth Disease (FMD) |
| title_full_unstemmed | Development and Evaluation of Molecular Pen-Side Assays without Prior RNA Extraction for Peste des Petits Ruminants (PPR) and Foot and Mouth Disease (FMD) |
| title_short | Development and Evaluation of Molecular Pen-Side Assays without Prior RNA Extraction for Peste des Petits Ruminants (PPR) and Foot and Mouth Disease (FMD) |
| title_sort | development and evaluation of molecular pen side assays without prior rna extraction for peste des petits ruminants ppr and foot and mouth disease fmd |
| topic | pen-side PCR peste des petits ruminants foot and mouth disease PPR FMD PCR |
| url | https://www.mdpi.com/1999-4915/14/4/835 |
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