Development of a Novel Tissue Blot Hybridization Chain Reaction for the Identification of Plant Viruses
Assays for the high throughput screening of crops for virus monitoring need to be quick, easy, and low cost. One method involves using tissue blot immunoassays (TBIA), where plant stems are blotted onto nitrocellulose membrane and screened with available antibodies against a range of viruses. TBIAs...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-09-01
|
| Series: | Plants |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2223-7747/11/17/2325 |
| _version_ | 1827665291857887232 |
|---|---|
| author | Fiona Filardo Peter Vukovic Murray Sharman Cherie Gambley Paul Campbell |
| author_facet | Fiona Filardo Peter Vukovic Murray Sharman Cherie Gambley Paul Campbell |
| author_sort | Fiona Filardo |
| collection | DOAJ |
| description | Assays for the high throughput screening of crops for virus monitoring need to be quick, easy, and low cost. One method involves using tissue blot immunoassays (TBIA), where plant stems are blotted onto nitrocellulose membrane and screened with available antibodies against a range of viruses. TBIAs are inexpensive but limited by antibody availability and specificity. To circumvent the antibody limitations, we developed the tissue blot hybridization chain reaction (TB-HCR). As with TBIA, plant stems are blotted onto a nitrocellulose membrane, however, TB-HCR involves using nucleic acid probes instead of antibodies. We demonstrated for the first time that TB-HCR can be used for plant viruses by designing and testing probes against species from several virus genera including <i>begomovirus</i>, <i>polerovirus</i>, <i>luteovirus</i>, <i>cucumovirus</i>, and <i>alfamovirus</i>. We also explored different hairpin reporter methods such as biotin/streptavidin-AP and the Alexa Fluor-488 Fluorophore. TB-HCR has applications for low-cost diagnostics for large sample numbers, rapid diagnostic deployment for new viruses, and can be performed as a preliminary triage assay prior to downstream applications. |
| first_indexed | 2024-03-10T01:23:18Z |
| format | Article |
| id | doaj.art-d2adc0df4a9248158479b99832e4128c |
| institution | Directory Open Access Journal |
| issn | 2223-7747 |
| language | English |
| last_indexed | 2024-03-10T01:23:18Z |
| publishDate | 2022-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Plants |
| spelling | doaj.art-d2adc0df4a9248158479b99832e4128c2023-11-23T13:56:47ZengMDPI AGPlants2223-77472022-09-011117232510.3390/plants11172325Development of a Novel Tissue Blot Hybridization Chain Reaction for the Identification of Plant VirusesFiona Filardo0Peter Vukovic1Murray Sharman2Cherie Gambley3Paul Campbell4Queensland Department of Agriculture and Fisheries, Ecosciences Precinct, GPO Box 267, Brisbane, QLD 4001, AustraliaQueensland Department of Agriculture and Fisheries, Ecosciences Precinct, GPO Box 267, Brisbane, QLD 4001, AustraliaQueensland Department of Agriculture and Fisheries, Ecosciences Precinct, GPO Box 267, Brisbane, QLD 4001, AustraliaQueensland Department of Agriculture and Fisheries, Ecosciences Precinct, GPO Box 267, Brisbane, QLD 4001, AustraliaQueensland Department of Agriculture and Fisheries, Ecosciences Precinct, GPO Box 267, Brisbane, QLD 4001, AustraliaAssays for the high throughput screening of crops for virus monitoring need to be quick, easy, and low cost. One method involves using tissue blot immunoassays (TBIA), where plant stems are blotted onto nitrocellulose membrane and screened with available antibodies against a range of viruses. TBIAs are inexpensive but limited by antibody availability and specificity. To circumvent the antibody limitations, we developed the tissue blot hybridization chain reaction (TB-HCR). As with TBIA, plant stems are blotted onto a nitrocellulose membrane, however, TB-HCR involves using nucleic acid probes instead of antibodies. We demonstrated for the first time that TB-HCR can be used for plant viruses by designing and testing probes against species from several virus genera including <i>begomovirus</i>, <i>polerovirus</i>, <i>luteovirus</i>, <i>cucumovirus</i>, and <i>alfamovirus</i>. We also explored different hairpin reporter methods such as biotin/streptavidin-AP and the Alexa Fluor-488 Fluorophore. TB-HCR has applications for low-cost diagnostics for large sample numbers, rapid diagnostic deployment for new viruses, and can be performed as a preliminary triage assay prior to downstream applications.https://www.mdpi.com/2223-7747/11/17/2325tissue blothybridization chain reactionself-assemblyplant virusesTYLCVpolerovirus |
| spellingShingle | Fiona Filardo Peter Vukovic Murray Sharman Cherie Gambley Paul Campbell Development of a Novel Tissue Blot Hybridization Chain Reaction for the Identification of Plant Viruses Plants tissue blot hybridization chain reaction self-assembly plant viruses TYLCV polerovirus |
| title | Development of a Novel Tissue Blot Hybridization Chain Reaction for the Identification of Plant Viruses |
| title_full | Development of a Novel Tissue Blot Hybridization Chain Reaction for the Identification of Plant Viruses |
| title_fullStr | Development of a Novel Tissue Blot Hybridization Chain Reaction for the Identification of Plant Viruses |
| title_full_unstemmed | Development of a Novel Tissue Blot Hybridization Chain Reaction for the Identification of Plant Viruses |
| title_short | Development of a Novel Tissue Blot Hybridization Chain Reaction for the Identification of Plant Viruses |
| title_sort | development of a novel tissue blot hybridization chain reaction for the identification of plant viruses |
| topic | tissue blot hybridization chain reaction self-assembly plant viruses TYLCV polerovirus |
| url | https://www.mdpi.com/2223-7747/11/17/2325 |
| work_keys_str_mv | AT fionafilardo developmentofanoveltissueblothybridizationchainreactionfortheidentificationofplantviruses AT petervukovic developmentofanoveltissueblothybridizationchainreactionfortheidentificationofplantviruses AT murraysharman developmentofanoveltissueblothybridizationchainreactionfortheidentificationofplantviruses AT cheriegambley developmentofanoveltissueblothybridizationchainreactionfortheidentificationofplantviruses AT paulcampbell developmentofanoveltissueblothybridizationchainreactionfortheidentificationofplantviruses |