Development and Validation of a New DIVA Real-Time PCR Allowing to Differentiate Wild-Type Lumpy Skin Disease Virus Strains, Including the Asian Recombinant Strains, from Neethling-Based Vaccine Strains
The current epidemic in Asia, driven by LSDV recombinants, poses difficulties to existing DIVA PCR tests, as these do not differentiate between homologous vaccine strains and the recombinant strains. We, therefore, developed and validated a new duplex real-time PCR capable of differentiating Neethli...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-03-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/15/4/870 |
| _version_ | 1797603254444490752 |
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| author | Andy Haegeman Ilse De Leeuw Wannes Philips Nick De Regge |
| author_facet | Andy Haegeman Ilse De Leeuw Wannes Philips Nick De Regge |
| author_sort | Andy Haegeman |
| collection | DOAJ |
| description | The current epidemic in Asia, driven by LSDV recombinants, poses difficulties to existing DIVA PCR tests, as these do not differentiate between homologous vaccine strains and the recombinant strains. We, therefore, developed and validated a new duplex real-time PCR capable of differentiating Neethling-based vaccine strains from classical and recombinant wild-type strains that are currently circulating in Asia. The DIVA potential of this new assay, seen in the in silico evaluation, was confirmed on samples from LSDV infected and vaccinated animals and on isolates of LSDV recombinants (n = 12), vaccine (n = 5), and classic wild-type strains (n = 6). No cross-reactivity or a-specificity with other capripox viruses was observed under field conditions in non-capripox viral stocks and negative animals. The high analytical sensitivity is translated into a high diagnostic specificity as more than 70 samples were all correctly detected with Ct values very similar to those of a published first-line pan capripox real-time PCR. Finally, the low inter- and intra-run variability observed shows that the new DIVA PCR is very robust which facilitates its implementation in the lab. All validation parameters that are mentioned above indicate the potential of the newly developed test as a promising diagnostic tool which could help to control the current LSDV epidemic in Asia. |
| first_indexed | 2024-03-11T04:27:35Z |
| format | Article |
| id | doaj.art-e8d11ab7c209433d981aec8836249a88 |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-11T04:27:35Z |
| publishDate | 2023-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-e8d11ab7c209433d981aec8836249a882023-11-17T21:44:57ZengMDPI AGViruses1999-49152023-03-0115487010.3390/v15040870Development and Validation of a New DIVA Real-Time PCR Allowing to Differentiate Wild-Type Lumpy Skin Disease Virus Strains, Including the Asian Recombinant Strains, from Neethling-Based Vaccine StrainsAndy Haegeman0Ilse De Leeuw1Wannes Philips2Nick De Regge3Sciensano, Infectious Diseases in Animals, Exotic and Vector-Borne Diseases, Groeselenberg 99, B-1180 Brussels, BelgiumSciensano, Infectious Diseases in Animals, Exotic and Vector-Borne Diseases, Groeselenberg 99, B-1180 Brussels, BelgiumSciensano, EURL for Diseases Caused by Capripox Viruses, Groeselenberg 99, B-1180 Brussels, BelgiumSciensano, Infectious Diseases in Animals, Exotic and Vector-Borne Diseases, Groeselenberg 99, B-1180 Brussels, BelgiumThe current epidemic in Asia, driven by LSDV recombinants, poses difficulties to existing DIVA PCR tests, as these do not differentiate between homologous vaccine strains and the recombinant strains. We, therefore, developed and validated a new duplex real-time PCR capable of differentiating Neethling-based vaccine strains from classical and recombinant wild-type strains that are currently circulating in Asia. The DIVA potential of this new assay, seen in the in silico evaluation, was confirmed on samples from LSDV infected and vaccinated animals and on isolates of LSDV recombinants (n = 12), vaccine (n = 5), and classic wild-type strains (n = 6). No cross-reactivity or a-specificity with other capripox viruses was observed under field conditions in non-capripox viral stocks and negative animals. The high analytical sensitivity is translated into a high diagnostic specificity as more than 70 samples were all correctly detected with Ct values very similar to those of a published first-line pan capripox real-time PCR. Finally, the low inter- and intra-run variability observed shows that the new DIVA PCR is very robust which facilitates its implementation in the lab. All validation parameters that are mentioned above indicate the potential of the newly developed test as a promising diagnostic tool which could help to control the current LSDV epidemic in Asia.https://www.mdpi.com/1999-4915/15/4/870real-time PCRlumpy skin diseaseDIVArecombinant strainvaccine strain |
| spellingShingle | Andy Haegeman Ilse De Leeuw Wannes Philips Nick De Regge Development and Validation of a New DIVA Real-Time PCR Allowing to Differentiate Wild-Type Lumpy Skin Disease Virus Strains, Including the Asian Recombinant Strains, from Neethling-Based Vaccine Strains Viruses real-time PCR lumpy skin disease DIVA recombinant strain vaccine strain |
| title | Development and Validation of a New DIVA Real-Time PCR Allowing to Differentiate Wild-Type Lumpy Skin Disease Virus Strains, Including the Asian Recombinant Strains, from Neethling-Based Vaccine Strains |
| title_full | Development and Validation of a New DIVA Real-Time PCR Allowing to Differentiate Wild-Type Lumpy Skin Disease Virus Strains, Including the Asian Recombinant Strains, from Neethling-Based Vaccine Strains |
| title_fullStr | Development and Validation of a New DIVA Real-Time PCR Allowing to Differentiate Wild-Type Lumpy Skin Disease Virus Strains, Including the Asian Recombinant Strains, from Neethling-Based Vaccine Strains |
| title_full_unstemmed | Development and Validation of a New DIVA Real-Time PCR Allowing to Differentiate Wild-Type Lumpy Skin Disease Virus Strains, Including the Asian Recombinant Strains, from Neethling-Based Vaccine Strains |
| title_short | Development and Validation of a New DIVA Real-Time PCR Allowing to Differentiate Wild-Type Lumpy Skin Disease Virus Strains, Including the Asian Recombinant Strains, from Neethling-Based Vaccine Strains |
| title_sort | development and validation of a new diva real time pcr allowing to differentiate wild type lumpy skin disease virus strains including the asian recombinant strains from neethling based vaccine strains |
| topic | real-time PCR lumpy skin disease DIVA recombinant strain vaccine strain |
| url | https://www.mdpi.com/1999-4915/15/4/870 |
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