Evaluation of Bluetongue Virus (BTV) Antibodies for the Immunohistochemical Detection of BTV and Other Orbiviruses
The detection of bluetongue virus (BTV) antigens in formalin-fixed tissues has been challenging; therefore, only a limited number of studies on suitable immunohistochemical approaches have been reported. This study details the successful application of antibodies for the immunohistochemical detectio...
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MDPI AG
2020-08-01
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| Series: | Microorganisms |
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| Online Access: | https://www.mdpi.com/2076-2607/8/8/1207 |
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| author | Fabian Z. X. Lean Jean Payne Jennifer Harper Joanne Devlin David T. Williams John Bingham |
| author_facet | Fabian Z. X. Lean Jean Payne Jennifer Harper Joanne Devlin David T. Williams John Bingham |
| author_sort | Fabian Z. X. Lean |
| collection | DOAJ |
| description | The detection of bluetongue virus (BTV) antigens in formalin-fixed tissues has been challenging; therefore, only a limited number of studies on suitable immunohistochemical approaches have been reported. This study details the successful application of antibodies for the immunohistochemical detection of BTV in BSR variant baby hamster kidney cells (BHK-BSR) and infected sheep lungs that were formalin-fixed and paraffin-embedded (FFPE). BTV reactive antibodies raised against non-structural (NS) proteins 1, 2, and 3/3a and viral structural protein 7 (VP7) were first evaluated on FFPE BTV-infected cell pellets for their ability to detect BTV serotype 1 (BTV-1). Antibodies that were successful in immunolabelling BTV-1 infected cell pellets were further tested, using similar methods, to determine their broader immunoreactivity against a diverse range of BTV and other orbiviruses. Antibodies specific for NS1, NS2, and NS3/3a were able to detect all BTV isolates tested, and the VP7 antibody cross-reacted with all BTV isolates, except BTV-15. The NS1 antibodies were BTV serogroup-specific, while the NS2, NS3/3a, and VP7 antibodies demonstrated immunologic cross-reactivity to related orbiviruses. These antibodies also detected viral antigens in BTV-3 infected sheep lung. This study demonstrates the utility of FFPE-infected cell pellets for the development and validation of BTV immunohistochemistry. |
| first_indexed | 2024-03-10T17:47:26Z |
| format | Article |
| id | doaj.art-f1e7369e04b44c96893f0821be6a69dd |
| institution | Directory Open Access Journal |
| issn | 2076-2607 |
| language | English |
| last_indexed | 2024-03-10T17:47:26Z |
| publishDate | 2020-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Microorganisms |
| spelling | doaj.art-f1e7369e04b44c96893f0821be6a69dd2023-11-20T09:29:38ZengMDPI AGMicroorganisms2076-26072020-08-0188120710.3390/microorganisms8081207Evaluation of Bluetongue Virus (BTV) Antibodies for the Immunohistochemical Detection of BTV and Other OrbivirusesFabian Z. X. Lean0Jean Payne1Jennifer Harper2Joanne Devlin3David T. Williams4John Bingham5CSIRO Australian Centre for Disease Preparedness (ACDP, formerly AAHL), Geelong 3220, Victoria, AustraliaCSIRO Australian Centre for Disease Preparedness (ACDP, formerly AAHL), Geelong 3220, Victoria, AustraliaCSIRO Australian Centre for Disease Preparedness (ACDP, formerly AAHL), Geelong 3220, Victoria, AustraliaDepartment of Veterinary Biosciences, Faculty of Veterinary and Agricultural Sciences, the University of Melbourne, Parkville 3052, Victoria, AustraliaCSIRO Australian Centre for Disease Preparedness (ACDP, formerly AAHL), Geelong 3220, Victoria, AustraliaCSIRO Australian Centre for Disease Preparedness (ACDP, formerly AAHL), Geelong 3220, Victoria, AustraliaThe detection of bluetongue virus (BTV) antigens in formalin-fixed tissues has been challenging; therefore, only a limited number of studies on suitable immunohistochemical approaches have been reported. This study details the successful application of antibodies for the immunohistochemical detection of BTV in BSR variant baby hamster kidney cells (BHK-BSR) and infected sheep lungs that were formalin-fixed and paraffin-embedded (FFPE). BTV reactive antibodies raised against non-structural (NS) proteins 1, 2, and 3/3a and viral structural protein 7 (VP7) were first evaluated on FFPE BTV-infected cell pellets for their ability to detect BTV serotype 1 (BTV-1). Antibodies that were successful in immunolabelling BTV-1 infected cell pellets were further tested, using similar methods, to determine their broader immunoreactivity against a diverse range of BTV and other orbiviruses. Antibodies specific for NS1, NS2, and NS3/3a were able to detect all BTV isolates tested, and the VP7 antibody cross-reacted with all BTV isolates, except BTV-15. The NS1 antibodies were BTV serogroup-specific, while the NS2, NS3/3a, and VP7 antibodies demonstrated immunologic cross-reactivity to related orbiviruses. These antibodies also detected viral antigens in BTV-3 infected sheep lung. This study demonstrates the utility of FFPE-infected cell pellets for the development and validation of BTV immunohistochemistry.https://www.mdpi.com/2076-2607/8/8/1207<i>Bluetongue virus</i>immunohistochemistry |
| spellingShingle | Fabian Z. X. Lean Jean Payne Jennifer Harper Joanne Devlin David T. Williams John Bingham Evaluation of Bluetongue Virus (BTV) Antibodies for the Immunohistochemical Detection of BTV and Other Orbiviruses Microorganisms <i>Bluetongue virus</i> immunohistochemistry |
| title | Evaluation of Bluetongue Virus (BTV) Antibodies for the Immunohistochemical Detection of BTV and Other Orbiviruses |
| title_full | Evaluation of Bluetongue Virus (BTV) Antibodies for the Immunohistochemical Detection of BTV and Other Orbiviruses |
| title_fullStr | Evaluation of Bluetongue Virus (BTV) Antibodies for the Immunohistochemical Detection of BTV and Other Orbiviruses |
| title_full_unstemmed | Evaluation of Bluetongue Virus (BTV) Antibodies for the Immunohistochemical Detection of BTV and Other Orbiviruses |
| title_short | Evaluation of Bluetongue Virus (BTV) Antibodies for the Immunohistochemical Detection of BTV and Other Orbiviruses |
| title_sort | evaluation of bluetongue virus btv antibodies for the immunohistochemical detection of btv and other orbiviruses |
| topic | <i>Bluetongue virus</i> immunohistochemistry |
| url | https://www.mdpi.com/2076-2607/8/8/1207 |
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