A simple and economical enzyme immunoassay technique using a polystyrene Petri dish as the solid support and a substrate mixed with agar to visualize enzymatic activity was developed and standardized for the detection of plant viruses. Reagents were added as drops into circular areas bounded with a hydrophobic cryomarker pen on the inner surface of the Petri dish. After subsequent incubation and washing steps with antiserum and alkaline phosphatase conjugate, the substrate (5-bromo-4-chloro-3-indolyl phosphate and nitroblue tetrazolium) solution was mixed with warm agar and added to the inner Petri dish surface. Distinct dark blue to purple dots were formed on the agar matrix at the sites where the virus was trapped. The technique is tentatively called 'Petri dish-Agar Dot Immunoenzymatic Assay' (PADIA). In addition to substituting the expensive microtitre plate or nitrocellulose membrane with a plastic Petri dish, PADIA consumes 5-10 times less reagents (antiserum and enzyme conjugate) than conventional ELISA or dot immunobinding assay (DIBA). The total assay cost was at least four times less than that of conventional ELISA. The technique was as sensitive as ELISA. PADIA can be used in poorly equipped laboratories with a minimum of input.
Petri dish-Agar Dot Immunoenzymatic Assay (PADIA) - a new and inexpensive method for the detection and identification of plant viruses
Reprints and Corporate Permissions
Please note: Selecting permissions does not provide access to the full text of the article, please see our help page How do I view content?
To request a reprint or corporate permissions for this article, please click on the relevant link below:
Academic Permissions
Please note: Selecting permissions does not provide access to the full text of the article, please see our help page How do I view content?
Obtain permissions instantly via Rightslink by clicking on the button below:
If you are unable to obtain permissions via Rightslink, please complete and submit this Permissions form. For more information, please visit our Permissions help page.
Related research
People also read lists articles that other readers of this article have read.
Recommended articles lists articles that we recommend and is powered by our AI driven recommendation engine.
Cited by lists all citing articles based on Crossref citations.
Articles with the Crossref icon will open in a new tab.