Symptoms of Grapevine red blotch-associated virus on a leaf of Vitis vinifera cv. Cabernet franc. Photo courtesy of Marc Fuchs, Cornell University
Agdia, Inc (Elkhart, IN) has commercialized a rapid and user-friendly DNA test kit, based on their AmplifyRP® Acceler8® platform, for detection Grapevine red blotch-associated virus (GRBaV) in grape.
GRBaV is an emerging threat to the grapevine industry. Depending on the grapevine cultivar, symptoms of the pathogen can vary. It can affect the profitability of vineyards by substantially reducing fruit quality and ripening according to an article in Phytopathology (Sudarshana et al, 2015).
Agdia’s AmplifyRP Acceler8 test is an isothermal DNA amplification assay based on recombinase polymerase amplification technology (RPA). RPA facilitates rapid nucleic acid (DNA or RNA) amplification at a single operating temperature of 39°C using crude sample extracts. RPA is different from laboratory based nucleic acid amplification and detection technologies, such as PCR, because it does not require thermal cycling or highly purified samples. RPA is also much faster than other methods because the amplification process is more efficient and is usually completed in 20 minutes or less.
To run the test an end-user would select tissue from a suspect plant and extract it in a provided buffer filled mesh-bag. They then add a small volume of the plant extract to a lyophilized reaction tube that has been reconstituted with a provided solution. The reaction is then heated to 39°C using a portable heat block (provided separately) for 20 minutes. During this time GRBaV DNA, if present, will be amplified to detectable levels. Once the amplification step is completed the end-user places the reaction into an amplicon detection chamber which reports results as either negative (one-line) or positive (two-lines) on a lateral flow strip contained inside the device. The total time to perform the test is about 45 minutes including the sample extraction step.
Test results from AmplifyRP Acceler8 for GRBaV. (Left = Positive GRBaV, Right = Negative)
Agdia claims the test is at least ten to one hundred times more sensitive than laboratory based PCR methods but can be completed in a fraction of the time by non-experienced users. The test was able to detect as few as 100 femtograms (fg) of total DNA purified from an infected GRBaV sample whereas PCR was able to detect only 10 picograms (pg) of the same purified DNA extract.
Agdia’s AmplifyRP Acceler8 kits for GRBaV come with everything needed to perform the test except for the portable heat block, pipette, and tips. The excluded items can be purchased separately in a starter pack for around $300. The cost of a kit is $240 and each kit contains 8 tests. The test is also offered as a service by Agdia Testing Services for those who wish to send samples for diagnosis. For more information on the test contact Agdia at 574-264-2014 or via email at info@agdia.com
About Agdia
A leading provider of diagnostic solutions for agriculture, Agdia, Inc. has been serving plant breeders, propagators, growers, universities, and private testing laboratories since 1981. The company offers a comprehensive portfolio of validated, easy-to-use diagnostics for identifying plant pathogens, hormones, and transgenic traits, as well as in-house testing services. Agdia is ISO 9001:2008 certified and their Testing Services Laboratory is ISO/IEC 17025:2005 accredited.
About AmplifyRP
AmplifyRP Acceler8 Test Kits employ recombinase polymerase amplification (RPA) technology licensed by Agdia, Inc (licensee) from TwistDx Limited, U.K (licensor). Use of the RPA process and probe technologies are protected by US patents 7,270,981 B2, 7,399,590 B2, 7,435,561 B2, 7,485,428 B2 and foreign equivalents in addition to pending patents.
AmplifyRP® and Acceler8® are registered trademarks of Agdia, Inc.
References
- Sudarshana MR, Perry KL, and Fuchs MF. 2015. Grapevine Red Blotch-Associated Virus, an Emerging Threat to the Grapevine Industry. Phytopathology.105:1026-32
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