Agdia Releases Rapid Isothermal Assay for Detection of Emerging Pathogen in Industrial Hemp

Agdia, Inc. (Elkhart, IN) is happy to announce the commercialization of a rapid, user-friendly, DNA-based assay, on their AmplifyRP® XRT platform, for the detection of Beet curly top virus.

Curly top disease affects numerous commercially important hosts, including common bean (Phaseolus vulgaris), cucumber (Cucumis sativus), industrial hemp (Cannabis sativa), pepper (Capsicum annuum), potato (Solanum tuberosum), spinach (Spinacia oleracea), squash and pumpkin (Cucurbita pepo), sugar and table beet (Beta vulgaris) and tomato (Solanum lycopersicum). This disease is caused by Beet curly top virus (BCTV), a Curtovirus (family Geminiviridae), existing as a complex of strains differentiated genotypically, causing symptomology on the hosts mentioned above. Additionally, strains of BCTV are known to infect more than 300 species of plants in no fewer than 44 families, many of which are asymptomatic, weedy hosts.

Symptomology of Beet curly top virus was first observed in the late 19th century in the western U.S. on sugar beets. It was, however, not recognized as being caused by a specific pathogen until 1915, when leafhopper transmission was proven, and viral etiology was proposed. Since then, BCTV has spread throughout North America where hosts are cultivated, including several states in the American West and Southwest, southwestern Canada and Mexico. Furthermore, BCTV has been identified in parts of South America and several countries in the Mediterranean basin. All strains of BCTV are considered pathogens of quarantine importance in Canada, Israel, Mexico and the European Union.

Beet curly top virus is transmitted efficiently by the beet leafhopper, Circulifer tenellus (Order Hemiptera), in a persistent circulative manner. The virus can be acquired within minutes of feeding, and insects are known to remain viruliferous for up to a month. Beet curly top virus is phloem-limited, and the leafhopper must feed on infected phloem to acquire and transmit the virus to healthy plants. Circulifer tenellus is the only known vector in North America; however, in Europe, C. opacipennis is also known to vector the virus. The robust dynamics of the host-virus-vector relationship facilitate epidemics in parts of the world where leafhopper populations are high. Furthermore, the movement of infected propagative materials can spread the virus across great distances. Mechanical transmission through infected plant sap has been accomplished under experimental conditions. Nevertheless, this scenario is not thought to contribute to the epidemiology of naturally occurring curly top infections. Seed transmission of BCTV is not known to occur in the host species listed above.

BCTV symptoms on tomato plants in outdoor cultivation
Figure 1: Beet curly top virus symptoms in outdoor tomato cultivation. Used with permission from Whitney Cranshaw, Colorado State University, Bugwood.org

Symptoms of curly top disease vary according to host and are typically more severe when plants are infected at earlier growth stages; many plants die before reaching maturity. Symptoms of curly top include stunted and distorted plant growth; leaf curling, crumpling, yellowing, vein swelling and distortion; and necrosis and hyperplasia of the phloem (figure 1). On beets, phloem tissue becomes necrotic, and exudate appears on the leaf surface. On tomato and pepper, fruit set is greatly diminished, and fruit that does form ripens prematurely. Furthermore, veins become purple.

Industrial hemp has reemerged as an important crop within several U.S. states, due to federal legalization and the demand for fiber, seed and cannabidiol. As production has increased, the list of disease organisms infecting this crop has grown to include several fungal, bacterial, viral and viroid pathogens. Beet curly top virus has been confirmed infecting industrial hemp and appears to be widespread on this crop throughout regions where vectors are present. There is a scarcity of research on this specific pathosystem; however, the understanding of the epidemiology of BCTV on cannabis is burgeoning, along with the crop. Symptoms of infection on industrial hemp include stunting leaf deformation and chlorosis (figure 2).

BCTV symptoms of leaf curling and chlorosis on cannabis plant
Figure 2: Beet curly top virus infection showing both yellowing and strong leaf curling in Cannabis spp. Used with permission from Whitney Cranshaw, Colorado State University, bugwood.org

Agdia’s new AmplifyRP® XRT assay for detection BCTV is based on recombinase polymerase amplification (RPA). This technology promotes the rapid amplification and detection of nucleic acid targets, DNA or RNA, while maintaining a single operating temperature of 39 – 42 °C. The AmplifyRP® XRT products achieve target sensitivity and specificity comparable to PCR, while having clear advantages over the lab-based technology. AmplifyRP® XRT products do not require a nucleic acid purification step; crude sample extracts are prepared using a simple extraction buffer and tested directly. This makes the testing process simple and saves the end user valuable time. Furthermore, this facilitates the implementation of this technology at remote locations with limited resources. When paired with Agdia’s AmpliFire® isothermal fluorometer (figure 3), the XRT system is a rapid, user-friendly tool that can be implemented in the field or the lab by personnel with limited experience in molecular diagnostics.

AmpliFire portable isothermal fluorometer with target and internal control curves on screen
Figure 3: AmpliFire portable isothermal fluorometer

Agdia states their assay was screened against a diverse collection of confirmed strains, including those infecting beets, industrial hemp, peppers and tomatoes, detecting all true positives. Furthermore, no cross-reactivity was observed with an extensive panel of viral and viroid pathogens, including Alfalfa mosaic virusCucumber mosaic virusHop latent viroidHop stunt viroidTobacco mosaic virusTobacco ringspot virusTomato brown rugose fruit virusTomato mosaic virusTomato ringspot virusTomato spotted wilt virus and Tomato yellow leaf curl virus. Sensitivity for this assay is greater than that observed with the published RT-qPCR assay and conventional RT-PCR assay to which it was compared. This product was developed to test leaf, stem and petiole tissue.

The introduction of this product brings Agdia’s catalog to 25 assays on the AmplifyRP® platform. High levels of market demand for field-deployable, plant pathogen detection products have driven this output, and Agdia maintains they will continue to expand their product offerings.

Agdia Now Accepting Hemp Samples for Pathogen Testing

Following decades of service to food crop and ornamental growers, Agdia’s in-house diagnostic laboratory is now offering its expansive disease testing program to the hemp market.

Deborah (Debi) Groth-Helms is the Director of Agdia’s Testing Services department. “We are excited to finally offer our comprehensive virus testing program to hemp growers and diagnosticians. For years we have worked with stock and seed producers in screening propagative tissues for the presence of key pathogens. Now we can help bolster the US hemp industry in their efforts to produce the healthiest plants possible.”

Agdia Testing Services offers multiple screening options and works with clients in customizing screens to best suit their needs. The basic Hemp Screen includes a panel of a dozen different tests for viruses, viroids, and other microorganisms.

According to Debi, “Information is being rapidly released on what causes disease in Cannabis. Having a big catalog of services already in place allows us to quickly respond to new reports. Experience working with closely related plant species helps us know what to monitor before an outbreak occurs.”

Included in the Hemp Screen are tests for Alfalfa mosaic virus (AMV), Cucumber mosaic virus (CMV), Hop latent viroid (HLVd), and many others. The Curto-Becurtovirus Group test listed in the screen detects all of the various virus species associated with Beet curly top disease. Additional tests are also available for less common or emerging pathogens. These highly sensitive molecular assays can be used to detect diseases such as witches’ broom (Phytoplasma Group PCR) and Lettuce chlorosis virus (Closteroviridae Group PCR).

Currently the laboratory is only accepting hemp tissues from US submitters that hold an active hemp license through the USDA, their state government, or tribal entity.  Information on hemp licensing and the 2018 Farm Bill can be found on the USDA’s website. The laboratory recommends shipping plant material with the United States Postal Service (USPS) along with a completed sample submission form.

Information on how to send samples can be found on the Agdia website. Questions regarding samples, customized screens, requests for quotes, and all other inquiries can be sent to info@agdia-emea.com

Agdia Commercializes ToBRFV ImmunoStrip® to Complete Diagnostic Suite

Agdia, Inc. (Elkhart, IN) has added an to their Tomato brown rugose fruit virus (ToBRFV) diagnostic suite. Their ImmunoStrip® for ToBRFV is the third diagnostic assay launched by Agdia in the first half of 2021 for detection of this pathogen. Their high-specificity ELISA assay for ToBRFV was released on January 7, followed by a rapid molecular AmplifyRP® XRT for ToBRFV on January 12.

YouTube Preview Image

Tomato brown rugose fruit virus is a resistance-breaking Tobamovirus that causes severe economic losses in solanaceous crops, including Solanum lycopersicum (tomato) and Capsicum spp. (pepper). It causes symptoms typical of Tobamoviruses that include mosaic and chlorosis on the leaves and discoloration and deformation of the fruit.  These symptoms decrease yield and render fruit unmarketable.

Tomato and pepper seeds, transplants and fruits from certain countries are subject to a USDA-APHIS Federal Import Order in the United States. Tomato brown rugose fruit virus has also been classified as a quarantine pathogen by EPPO (European and Mediterranean Plant Protection Organization).

Agdia’s ImmunoStrip® for ToBRFV has been validated for use with tomato, pepper and petunia samples and has been tested against ToBRFV isolates from around the globe.

Cross-reactivity was observed with a Tomato mottle mosaic virus (ToMMV) sample. Very mild cross-reactivity was observed with Tobacco mosaic virus (TMV) and Tomato mosaic virus (ToMV) samples. No other cross-reactions were observed when testing other Tobamoviruses, including Bell pepper mosaic virus (BPeMV), Cucumber green mottle mosaic virus (CGMMV), Kyuri green mottle mosaic virus (KGMMV), Pepper mild mottle virus (PMMoV), Tobacco mild green mosaic virus (TMGMV), and Zucchini Green Mottle Mosaic Virus (ZGMMV).

Agdia’s ImmunoStrip® platform provides end-users with a high level of utility. ImmunoStrips® are fit for use in the lab or in the field by those having no previous diagnostic experience, and results are visualized within 30 minutes.  Increasing levels of market demand persists for field-deployable plant pathogen detection products and Agdia maintains they will continue to expand this area of their catalog.

Agdia Adds to Diagnostic Suite for ToBRFV with Release of a Rapid, RNA-based Isothermal Molecular Kit

Agdia, Inc. (Elkhart, IN) has added another test kit to their Tomato brown rugose fruit virus (ToBRFV) diagnostic suite. AmplifyRP® XRT for ToBRFV is the second of three ToBRFV diagnostic assays to be launched by Agdia in the first half of 2021. Their high-specificity ELISA assay for ToBRFV was released on January 7. An ImmunoStrip® lateral flow device for ToBRFV is expected to be released in the coming months (Q1 or Q2 of 2021.)

Agdia’s AmplifyRP® XRT for ToBRFV has been validated for use with tomato and pepper seeds and leaf. As a rapid, field-deployable molecular method requiring far less training than traditional PCR methods, this assay provides users with greater flexibility to deploy detection capabilities where they need it, when they need it. Use cases for this assay include:

  • Laboratory-based molecular diagnosis with crude or purified extracts with faster time-to-result than traditional PCR or qPCR methods. This assay can be used with Agdia’s AmpliFire® isothermal fluorometer or with most commonly-used real-time PCR machines.
  • In-field monitoring at remote production sites as a stand-alone assay.
  • Directly test ELISA extracts for molecular confirmation of serological screening results, allowing you to act immediately to address production issues.

Agdia’s AmplifyRP® XRT for ToBRFV is highly specific to ToBRFV and has been experimentally proven to detect isolates from around the world. No cross-reactivity was observed with high titer samples from other Tobamoviruses, including Cucumber green mottle mosaic virus (CGMMV), Kyuri green mottle mosaic virus (KGMMV), Pepper mild mottle virus (PMMoV), Tobacco mosaic virus (TMV), Tomato mosaic virus (ToMV), Tobacco mild green mosaic virus (TMGMV), Zucchini Green Mottle Mosaic Virus (ZGMMV) and more.

Tomato brown rugose fruit virus is a resistance-breaking Tobamovirus that causes severe economic losses in solanaceous crops, including Solanum lycopersicum (tomato) and Capsicum spp. (pepper). It causes symptoms typical of Tobamoviruses that include mosaic and chlorosis on the leaves and discoloration and deformation of the fruit.  These symptoms decrease yield and render fruit unmarketable.

Tomato and pepper seeds, transplants and fruits from certain countries are subject to a USDA-APHIS Federal Import Order in the United States. Tomato brown rugose fruit virus has also been classified as a quarantine pathogen by EPPO (European and Mediterranean Plant Protection Organization).

A DNA RAPID TEST FOR THE DETECTION OF XYLELLA FASTIDIOSA

Agdia-Emea has commercialized a rapid and user-friendly DNA test kit, based on the AmplifyRP® XRT+ platform, for detection of Xylella fastidiosa (Xf).

Olive tree infected by Xylella Fastidiosa

Olive tree infected by Xylella Fastidiosa

Xylella fastidiosa (Xf) is the causal agent of multiple crop diseases including Pierce’s disease in grape, citrus variegated chlorosis (CVC), almond leaf scorch disease and olive quick decline syndrome. Severe Xf infections can decimate entire crops and cause enormous financial losses to growers.
The test is a real-time isothermal DNA amplification assay that utilizes recombinase polymerase amplification (RPA) technology.  RPA facilitates rapid nucleic acid (DNA or RNA) amplification at a single operating temperature of 39°C using crude sample extracts.   RPA can provide the same level of sensitivity and specificity as laboratory based PCR methods, but offers some clear advantages.  The total assay time is less than 30 minutes, including sample preparation.   This is a stark contrast to PCR methods which can take hours or days to complete.  PCR analysis must also be completed in a laboratory whereas AmplifyRP XRT+ can be performed on-site.

Agdia-Emea AmplifyRP XRT+ platform works similarly to the recently launched XRT platform in that the results can be obtained using a portable fluorometer.  However, XRT+ provides greater flexibility because the test can alternatively be performed using a small portable heat block with results being visualized post-amplification on an Amplicon Detection Chamber.  This allows end-users who don’t want to invest in a fluorometer an alternative method of running the test.

Photo 1: Real-time results on AmpliFire®

Photo 1: Real-time results on AmpliFire®

To run the test, an end-user first extracts their sample by adding it, along with extraction buffer, to a micro centrifuge tube that is provided in the kit.  After the sample has been extracted a small volume is added to a solution that is used to rehydrate a lyophilized reaction pellet.  Once the reaction pellet has been rehydrated it can be added to a portable fluorometer or a small heat block for 20 minutes.  During this time, if present, Xf DNA will be amplified.  If using the fluorometer results will automatically be displayed in real-time (photo 1).  Alternatively, if using a heat block, the reaction is added to an Amplicon Detection Chamber following the amplification step where results will be visible on a lateral flow strip (photo 2). 

Photo 2: Portable heat-block with post-amplification detection using Amplicon Detection Chambers

Photo 2: Portable heat-block with post-amplification detection using Amplicon Detection Chambers

The kit includes 24 or 48 reactions and the necessary buffers to perform the test. If performing the test as a real-time assay, the end-user will need to purchase a portable fluorometer separately.  Agdia claims the test is compatible with several commercially available fluorometers but recommends the AmpliFire® (Photo 1), manufactured by Douglas Scientific.  The AmpliFire is battery operated and has a small footprint, making it possible to test in remote locations. 

Alternatively, if the end-user runs the test as an end-point detection assay they will need to purchase Amplicon detection chambers, as well as a starter pack, separately.   The starter pack includes a heat block (necessary for the amplification step) as well as pipettes, pipette tips, and a tube rack.

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-Emea at +33 1 60 78 81 64 or via email at info@agdia-emea.com

Agdia Launches a 30-minute DNA test for Clavibacter detection in tomato

Agdia, Inc (Elkhart, IN) has developed a rapid and user-friendly DNA test kit, based on their AmplifyRP® XRT platform, an isothermal amplification method, for detection of Clavibacter michiganensis subsp. michiganensis (Cmm) in tomato.

Cmm, the causal agent of bacterial canker of tomato, is a devastating disease that can result in crop losses of up to 100% if not quickly identified and managed. Agdia’s AmplifyRP XRT test for Cmm provides growers and diagnostic laboratories a new and more powerful tool to assist in the diagnosis of this important quarantine pathogen.

The test is a real-time isothermal DNA amplification assay that utilizes recombinase polymerase amplification (RPA) technology.  RPA facilitates rapid nucleic acid (DNA or RNA) amplification at a single operating temperature of 39°C using crude sample extracts.   RPA can provide the same level of sensitivity and specificity as laboratory based PCR methods, but offers some clear advantages.  The total assay time is less than 30 minutes, including sample preparation.   This is a stark contrast to PCR methods which can take hours to complete.  PCR analysis must also be completed in a laboratory whereas AmplifyRP XRT can be performed on-site using a battery operated portable fluorometer.  Agdia states their new test is capable of detecting concentrations of Cmm bacteria as low as 200 cfu/mL, which offers greater sensitivity than any other commercially available test kit.

Results displayed on AmpliFire®

Results displayed on AmpliFire®

To run the test, an end-user first extracts their sample using a mesh extraction bag.  After the sample has been extracted a small volume is added to a solution that is used to rehydrate a lyophilized reaction pellet.  Once the reaction pellet has been rehydrated, it is added to a portable fluorometer for 20 minutes.  During this time Cmm DNA, if present, will be amplified and will produce a fluorescent signal that is detected by the instrument in real-time. 
Agdia claims the test is compatible with several commercially available fluorometers but recommends the AmpliFire® manufactured by Douglas Scientific.  The AmpliFire® is battery operated and has a small footprint, making it possible to test in remote locations.  Setup is simple because all the assay parameters are loaded into the AmpliFire® using a barcode that is affixed to the tests packaging.  It also has a large, intuitive, touch screen interface that is used to initiate reactions and view results.   

The test is also offered as a service by Agdia Testing Services for those who wish to send samples for diagnosis. 

For more information about the test please contact us:

Phone +33 (0) 1 60 78 81 64
Email : info@agdia-emea.com

AmplifyRP® is a registered Trademark of Agdia, Inc.

AmpliFire® is a registered Trademark of Douglas Scientific.   

Challenges of Phytopathologists working in the Seed Industry workshop

  • Agdia-Emea w

CLICK HERE FOR REGISTRATION

  • Subject: Challenges of Phytopathologists working in the Seed Industry                    
  • Date: Wednesday 22 March 2017
  • Time: 9.00 – 17.30
  • Location: Naktuinbouw, Sotaweg 22, Roelofarendsveen

New CGMMV ELISA test

We have launched a new ELISA test for Cucumber green mottle mosaic virus (CGMMV) that offers improved sensitivity.  Agdia’s new ELISA offers more than a two-fold increase in sensitivity compared to the prior version and it does not cross-react with other tobamoviruses.

CGMMV is a Tobamovirus that is transmitted mechanically and through infected seed in cucurbit species.  It is a major concern to commercial growers as well as seed producers because of its ability to cause severe reduction in crop quality, yield and profitability.  The virus was first reported on melon in the United States in 20131 and is considered a quarantined pathogen by USDA APHIS. 

The CGMMV ELISA is sold as a reagent set in 96, 500, 1000, and 5000 test well sizes.  Agdia provides a 1 year warranty on purchased reagents. For more information on the CGMMV ELISA e-mail info@agdia-emea.com, phone +33 1 60 78 81 64.

References:

1T. Tian and K. Posis, Plant Pest Diagnostics Center, California Department of Food and Agriculture (CDFA), Sacramento 95832; C. J. Maroon-Lango, V. Mavrodieva and S. Haymes, USDA APHIS (PPQ), BARC-East, Beltsville, MD 20705; and T. L. Pitman and B. W. Falk, Department of Plant Pathology, University of California, Davis 95616

New Four-Trait ImmunoStrip® for Testing Bollgard II® XtendFlex® Cotton

Agdia, Inc (Elkhart, IN) has announced the release of a new ImmunoStrip for detection of DMO, CP4 EPSPS, Bt-Cry2Ab, and Bt-Cry1Ac proteins in cotton seed and leaves.

Extremely fast and easy to use, this new ImmunoStrip is specially designed for single seed and single leaf testing and typically provides results within 3 to 5

 

minutes

after sample preparation.  Four independent test lines on the ImmunoStrip clearly identify and discriminate the target proteins when present in the sample.

The Agdia DMO, CP4 EPSPS, Bt-Cry2Ab, and Bt-Cry1Ac ImmunoStrip is available in 50-count packages. ImmunoCombs® are also available for high-throughput testing in 48- plate formats. As with all Agdia agricultural diagnostic products, the four-trait ImmunoStrip is backed by an extremely knowledgeable technical support team.

To order or learn more about Agdia’s four-trait ImmunoStrip for DMO, CP4 EPSPS, Bt-Cry2Ab, and Bt-Cry1Ac e-mail us at info@agdia-emea.com, phone +33 1 60 78 81 64

Bollgard®, Bollgard II®, Roundup Ready® Flex, and XtendFlex® are registered trademarks of Monsanto Technology LLC.

New Four-Trait ImmunoStrip® for Testing Bollgard II® XtendFlex® Cotton

We announce the release of a new ImmunoStrip for detection of DMO, CP4 EPSPS, Bt-Cry2Ab, and Bt-Cry1Ac proteins in cotton seed and leaves.
Extremely fast and easy to use, this new ImmunoStrip is specially designed for single seed and single leaf testing and typically provides results within 3 to 5 minutes after sample preparation.  Four independent test lines on the ImmunoStrip clearly identify and discriminate the target proteins when present in the sample.
The Agdia DMO, CP4 EPSPS, Bt-Cry2Ab, and Bt-Cry1Ac ImmunoStrip is available in 50-count packages. ImmunoCombs® are also available for high-throughput testing in 48- plate formats. As with all Agdia agricultural diagnostic products, the four-trait ImmunoStrip is backed by an extremely knowledgeable technical support team.
To order or learn more about Agdia’s four-trait ImmunoStrip for DMO, CP4 EPSPS, Bt-Cry2Ab, and Bt-Cry1Ac e-mail us at info@agdia-emea.com, phone +33 1 60 78 81 64

Bollgard®, Bollgard II®, Roundup Ready® Flex, and XtendFlex® are registered trademarks of Monsanto Technology LLC.

Agdia Accredited Under New USDA APHIS Testing Pilot Program for Minituber Testing

Agdia, Inc. (Elkhart, IN) has announced that their Testing Services department is the first private laboratory to be accredited under a new USDA APHIS Minituber Pathogen Testing Pilot program. This recent recognition by the USDA of Agdia Testing Services as an APHIS approved Testing Laboratory enables results of Testing Activities to be used to apply for minituber phytosanitary certification.

Agdia’s ISO/IEC 17025:2005 accredited Testing Services laboratory has conducted diagnostic testing for potato pathogens for 35 years and offers a full range of testing options for a multitude of crops.  Most recently, Agdia announced the implementation of a PCR method for detection of Dickeya spp., bacteria that have become a significant concern of U.S. potato producers in recent years.  Agdia Testing Services also routinely tests for the presence of Bacterial Ring Rot (BRR), a zero tolerance pathogen in seed certification, as well as several potato viruses such as Potato virus Y.  A complete list of tests that Agdia performs for potato pathogens can be found on their website.

 “We are grateful for the opportunity to work with the USDA in this new program for potato export. I am excited to see our laboratory recognized for the quality and breadth of our services. This new pilot program will help streamline communication with foreign government agencies importing US minitubers and will help the industry define what the most pressing disease concerns are for potato producers worldwide ”  says Deborah Groth-Helms, Director of Agdia Testing Services.

For more information on Agdia’s Testing Services laboratory please contact us at info@agdia-emea.com or
+33 1 60 78 81 64.

More information about the USDA APHIS Minituber Pathogen Testing Pilot program can be found by following the link below. 

https://www.aphis.usda.gov/aphis/ourfocus/planthealth/sa_export/sa_acns/ct_accreditation

Agdia Announces Implementation of DNA Test for Dickeya Spp. in Potato

Agdia, Inc. (Elkhart, IN) has announced the implementation of a new PCR testing service for diagnosis of Dickeya spp. in potato.  

lg_04719362-5056-A015-DA9AC54907244D93-1

Blackleg symptoms at base of stem. Photo courtesy of Amy Charkowski, Colorado State University.

Dickeya species of bacteria that cause blackleg disease, are pathogens of specific concern for American potato producers.  Dickeya dianthicola and Dickeya solani have caused significant crop losses in Northern Europe for the past decade and recently, Dickeya was found on the Eastern seaboard of the U.S. Since their discovery, these pathogens have been identified in other states in the Mid-Atlantic and Midwest growing regions and most recently Ontario, Canada.  Dickeya species are primarily spread through the planting of infected seed potato and favor warm and wet climates.  Symptoms may not be present on infected tubers and the disease may not manifest itself until after planting.  Buying certified seed lots that have been tested for Dickeya is crucial to preventing the spread of the pathogen. 

Agdia has recently validated a PCR testing protocol for detection of Dickeya.  PCR is a highly sensitive testing method used for amplifying pieces of DNA that are unique to a specific target of interest, in this case the member species of the genus Dickeya.  Using this method, Agdia Testing Services can reliably detect 1 infected tuber core in a composite of 100 cores.  By request, the same sample can be tested for bacterial ring rot (Cms), another bacteria of significant concern to potato growers.  Agdia offers reduced pricing when combining these two tests for the same submission of potato tubers, stems, or plantlets.

“The release of this new, validated testing service comes at a critical time for our potato growers worldwide. Routine PCR testing of production materials prior to distribution is the only way to control the spread of Dickeya. We hope to work with the USDA to ensure that U.S. potatoes are certified for export, and we will continue to test samples from international production locations to help control the spread of this disease and others” says Deborah Groth-Helms, Director of Agdia Testing Services.

Agdia’s Testing Services laboratory is an ISO/IEC 17025:2005 accredited laboratory and has recently obtained accreditation under a new USDA APHIS Potato Minituber Pathogen Testing Pilot program.  Under this pilot program, Agdia’s testing reports may be used as official documents for issuance of phytosanitary certificates related to export of U.S. potato minitubers.

New ImmunoStrip® for Detection of Tomato leaf curl New Delhi virus (ToLCNDV)

IMG_0415We are pleased to announce the immediate availability of a new rapid ImmunoStrip test for field detection of Tomato leaf curl New Delhi virus (ToLCNDV).

ToLCNDV is a whitefly transmitted begomovirus that is known to infect important solanaceous crops like tomato and pepper.  It also has been reported in cucurbits and melons.    According to an EPPO (European and Mediteranean Plant Protection Organization) bulletin it has become a widespread problem in parts of Asia and Europe and can cause severe symptoms in infected crops and heavy economic losses.
Agdia’s new ImmunoStrip is designed to allow onsite diagnosis of ToLCNDV in minutes.  This is particularly useful for a grower or a field inspector because the results are acquired in real-time and can help facilitate quicker management decisions.

To use the test the end-user only needs to take a sample from a suspicious plant, insert it into a buffer filled extraction bag and grind it up using a pen or marker.  Once the sample has been extracted, which takes only a minute or two, the ImmunoStrip is added to the extract and begins to work.  Positive results are typically visible in 5 to 10 minutes.  Agdia recommends running the test for 30 minutes to rule out weak infections where lower concentrations of the virus may exist.
Agdia-Emea offers ToLCNDV ImmunoStrip kits in sizes of 5 and 25 strips.  Test kits include all the necessary materials to perform the assay.  For more information, Agdia-Emea can be contacted by phone at +33 1 60 78 81 64or by email at info@agdia-emea.com

New rapid molecular detection test for Tomato chlorotic dwarf viroid

Tomato chlorotic dwarf viroid (TCDVd) is a serious problem in tomato crops due to its ability to reduce yield and produce unmarketable fruit.  Solanaceous ornamental crops that harbor the pathogen can serve as an inoculum source for the pathogen which can be transmitted to tomato.  Routine testing for the pathogen is useful to help identify the pathogen and prevent its spread.
Viroids are unique from plant viruses because they are naked small RNAs that do not code for a coat protein.  Immunoassays that are used in most plant virus testing applications are not an option.  The lack of a TCDVd coat protein forces a molecular detection approach to be used to target TCDVd RNA.  Typically this is done in a laboratory, using a technique known as polymerase chain reaction (PCR), by highly skilled technicians using sophisticated and expensive equipment. 
Amplify_RP Acceler8Agdia’s AmplifyRP Acceler8 test for TCDVd is the first commercially available test to allow producers to test for TCDVd themselves on-site. It works by using an isothermal amplification technology called recombinase polymerase amplification (RPA).  This technology allows rapid DNA or RNA amplification at constant operating temperature using a crude plant extract.
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 TCDVd RNA, 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.

The tests sensitivity is approximately 100 fg of in vitro transcribed TCDVd or 400 pg of the total plant RNA per reaction.  It is specific to TCDVd and does not cross-react with other Pospiviroids.
Agdia’s AmplifyRP Acceler8 kits for TCDVd 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. 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 please contact Agdia-Emea at +33 1 60 78 81 64 or via email at info@agdia-emea.com

Agdia Owners Named 2016 Indiana Small Business Persons of the Year by SBA

Agdia, Inc (Elkhart, IN) co-owners Baziel Vrient and Albert Vrient have been named the 2016 Indiana Small Business Persons of the Year by the U.S. Small Business Association (SBA).

Albert Vrient (left) and Baziel Vrient (right)

Albert Vrient (left) and Baziel Vrient (right)

The SBA annually awards a “Small Business Person(s) of the Year” from each of the 50 states, the District of Columbia, Puerto Rico, the U.S. Virgin Islands, and Guam.  The nominees are selected based on a number of criteria that include the company’s history of growth in revenue and staff, the innovativeness of their products and / or services, as well as their contributions to the community.
Agdia was founded in 1981 and is entering its 35th year in business.  They serve more than 150 countries with diagnostic solutions aimed at detecting plant pathogens and specific proteins that are produced by transgenic (GMO) crops.  “Agdia has always been focused on innovation and identifying ways to make testing accessible to any type of end-user, not just laboratory people.  Plant disease is responsible for billions of dollars of loss in the agricultural every year.  The more tools people have to prevent and tackle disease issues, the better chance they have to grow higher quality crops” says Baziel Vrient, Agdia’s President.  
Vrient went on to say that “Albert and I are proud to receive this award for Agdia as well as for the state of Indiana.  But more importantly I want to recognize all of our employees that make this business happen on a day to day basis as well as Chet and Jane Sutula for founding this business that provides a livelihood for 60 people and touches millions of lives on a day to day basis.  Also a special thanks goes out to Dana Trowbridge, Vice President of Business Banking for the First State Bank of Middlebury as well as Dan Jones, President of the Business Development Corp for nominating us for the award as well as having a firm belief in Agdia and the value we bring to the agricultural industry.”

Albert Vrient, Operations Manager of Agdia, also stated “We are excited to receive the recognition from the SBA and to be selected as Indiana’s Small Business Persons of the year.  This is a humbling award that would not be possible without having an elite team beside us everyday striving to offer the best possible products and services to the agricultural industry.  This award is as much theirs as it is ours.”  

The father-son team are now eligible to be selected as the National Small Business Persons of the year during National Small Business week in Washington D.C.  The winner will be selected the first week of May.

Agdia-Emea sponsor of the 13th International Plant Virus Epidemiology Symposium

Agdia-Emea is pleased to announce the official sponsorship of the 13th International Plant Virus Epidemiology Symposium, scheduled from Monday 6 June – Friday 10 June 2016 in Avignon, France.

This symposium will be a forum to discuss the latest advances in all areas of plant virus epidemiology and will stimulate exchanges between the actors involved in the (re)search of integrated and sustainable ways to manage virus diseases.

For more information on the symposium program please click here:

logo-partenaire_inra_logo

New ELISA and Flashkit® tests for testing eCry3.1Ab protein expressed in Agrisure® Duracade® corn

We are pleased to announce the commercialization of a new ELISA and lateral flow test strip which was developed to detect eCry3.1Ab protein expressed in transgenic corn, event 5307.

The eCry3.1Ab ELISA was specifically developed and validated for trait confirmation testing of seed lots using either individual kernels or leaf material. The assay is sensitive enough to detect 1 eCry3.1Ab kernel in 100 negative kernels (1%).

The Agdia eCry3.1Ab ELISA is a complete kit, including antibody-coated microtiter plates, enzyme conjugate, positive control, and extraction/wash buffers. Consistent with other ELISA assays in Agdia’s trait testing portfolio, the eCry3.1Ab protocol was designed with high-throughput testing and ease-of-use in mind. Utilizing a common extraction process and short incubation steps the entire test is completed in less than 2 hours.

The eCry3.1Ab FLASHKIT was specifically developed and validated for trait confirmation testing of seed lots using either individual kernels or leaf material. The ImmunoStrip may also be used for testing bulk grain samples with a detection limit of 1 eCry3.1Ab kernel in 200 negative kernels (0.5%). All Agdia ImmunoStrips are easy to use and are rugged enough for field environments and results are available in as little as 5 minutes.

Participation to EPPO Conference on Diagnostics for Plant Pests

EPPO Conference Dec2015
We had the pleasure to take part of the EPPO (European and Mediterranean Plant Protection Organization) Conference held in Angers, France in 1-2 December 2015, on diagnostics of plant pests.
This conference gave the opportunity to have several presentations on new molecular and bio-analytical identification tools, new tools for better morphological identification, improved approaches for early detection and other topics of importance in diagnostics.

EPPO Conference Dec2015

Experts have been invited also to know more about Agdia current R&D projects and new products available as the platform AmplifyRP, an isothermal amplification technique helping in plant pathogen detection.

For more information on this conference please click here: EPPO Conference on Diagnostics for Plant Pests

Agdia-Emea participation to USAID MLN Project Workshop at Nairobi

We are pleased to announce our participation as speakers to the USAID-funded MLN (Maize Lethal Necrosis) Project Inception Workshop (Nov 10, 2015) at Nairobi, Kenya.
USAID has recently approved a Project led by CIMMYT on Controlling the spread and impact of MLN in Sub-Saharan Africa through improved diagnostic capacity and MCMV-free commercial seed production.

Agdia-Emea and Agdia Inc. will present the MLN detection tool box including ImmunoStrip (Flashkit) quick tests and ELISA kits for MCMV and other viruses involved in the disease development.

We look forward to interact with the stakeholders participating in this meeting, which will include scientists from public and private sector institutions, representatives of National Plant Protection Organizations (NPPOs) and commercial seed sector, and USAID.

Detection of Maize lethal necrosis disease in the field now possible

MCMV ImmunoStripMaize lethal necrosis (MLN) is a severe disease affecting corn. The disease is caused by the combination of Maize chlorotic mottle virus (MCMV) and a virus belonging to the potyviridae family, commonly Sugarcane Mosaic Virus (SCMV). The viruses are vectored by insects as maize thrips, rootworms and leaf beetles.
Corn is one of the main staple foods in East Africa, consequently the Maize Lethal Necrosis Disease is considered as a serious threat for food security. Moreover, corn plants infected with MLN can develop secondary fungal infections and are not considered safe to eat. The disease has been first described in Kenya in 2011 (Wangai et al., 2012) and it has now spread to Uganda, Rwanda, Tanzania and South Sudan often with devastating consequences for corn fields.
If not controlled effectively, MLN can cause total crop loss. Since years a lot of efforts were made by different institutes to identify and manage the disease. The first step in disease control and management is to have rapid, accurate and cost effective diagnostic methods allowing to detect the viruses in the seeds before sowing and to detect the symptoms in the field at very early stages.

Agdia-Emea diagnostic kits and products are routinely used by many laboratories in several East African countries.
To help the scientists and farmers to fight against MLN, our company has recently developed a rapid kit, the MCMV and Potyvirus Immunostrip, allowing rapid and easy disease detection the fields. Thanks to ImmunoStrips® it will be possible to detect immediately the viruses causing MLN, as the first symptoms appear, without the need to ship the samples to a laboratory. This will allow a better monitoring of the disease and help the fight against the disease spread. The test requires no technical training and it is very simple to use. The answer is provided by the test is 30 minutes or less.

Please contact us for more information: info@agdia-emea.com

Reference: Wangai A et al. (2012) First Report of Maize chlorotic mottle virus and Maize Lethal Necrosis in Kenya. Plant Disease 96: 1582.

AGDIA receives ISO 9001:2008 Certification and ISO/IEC 17025:2005 Accreditation

ChrysanthemeWe have the pleasure to announce that Agdia, Inc. (Elkhart, IN) received both ISO 9001:2008 certification and ISO/IEC 17025:2005 accreditation.
Agdia obtained the ISO 9001:2008 certification from Smithers Quality Assessments. Being certified to this standard confirms that Agdia has implemented and follows a quality management system that focuses on continuously improving product quality and customer satisfaction. Agdia’s quality management system also ensures that corrective actions are taken when a defect in product quality is identified.
 
Agdia’s ISO/IEC 17025 accreditation was received from Laboratory Accreditation Bureau (L-A-B). This accreditation certifies that Agdia has the technical competence to a defined “Scope of Accreditation” and operates their laboratory according to a defined quality management system. Agdia’s scope of accreditation encompasses serological and molecular testing of plants for plant pathogens which includes testing methods such as ELISA, ImmunoStrip, PCR and real-time PCR.
 
Agdia’s decision to be audited and registered to both Standards was driven by the fact that they are both a diagnostic kit manufacturer as well as a diagnostic laboratory for the agricultural industry. Baziel Vrient, President of Agdia, stated that “Agdia has been committed to providing quality products and services to our customers since we were founded in 1981. As we grow, we must continuously evaluate our quality management systems and improve them to ensure we meet the evolving demands of our customers. This achievement is a testament to this commitment that we hold so dear.”

New High Sensitivity ELISA to Test For MCMV Associated With Maize Lethal Necrosis Disease

Vip3A corn

We have the pleasure to announce the commercialization of a new Enzyme-linked Immunosorbent Assay (ELISA) for detection of Maize chlorotic mottle virus (MCMV) in corn.

MCMV is of particular concern to corn producers because of its co-contribution to the devastating Maize Lethal Necrosis Disease (MLND). MLND can manifest itself when a crop is co-infected with MCMV and a member of the Potyvirus group, such as Sugarcane mosaic virus. African corn growing regions, where food production is vital, have been devastated by the disease in recent years.
Agdia’s new ELISA test for MCMV has been successfully validated against a global collection of isolates and is claimed to be a minimum of ten-fold more sensitive than any other serological method currently available. Agdia has initially released the test for foliar tissue testing but will soon be validating a protocol for testing bulk seed samples. Agdia states the MCMV seed protocol will be harmonized with methods currently in use by industry for other pathogens such as Pantoea stewartii (A.K.A. Stewart’s wilt / leaf blight of Corn).
MCMV ELISA reagents are immediately available in sizes of 96, 500, 1000, and 5000 testwells and can be ordered from Agdia-Emea using product number SRA 17002. Agdia Testing Services can also perform laboratory testing using the new test for producers who need to send their samples for third-party analysis. Agdia has later plans to commercialize a field-based Flashkit® (ImmunoStrip®) test for detection of MCMV this summer.

For more information please contact us: info@agdia-emea.com

New Molecular Hybridization Assays for Begomovirus and Curtovirus/Becurtovirus

Nucleic acid hybridization (NAK) also known as dot-blot hybridization has been used since the 1980s with increased specificity compared to ELISA. Agdia has enhanced the assay by developing a new buffer, AG1 Lysis Buffer, which simplifies the extraction process compared to traditional nucleic acid extraction techniques. These tests utilize multiple nucleic acid probes, which are labeled to target complimentary single-stranded nucleic acid on a nylon membrane.
Getting results is as easy as preparing the samples and spotting them onto the supplied membrane, where the targeted nucleic acids become immobilized. Once the step has been completed, the end-user mails the membrane to Agdia Testing Services for processing. Agdia’s laboratory will then introduce the membrane to labeled probes where it will hybridize to complementary nucleic acid structures, if present. Finally, chemiluminescent detection is used to produce a visual spot by exposing the membrane to film, indicating a positive result.

Molecular Hybridization membraneAgdia has released, with immediate availability, the following nucleic acid hybridization assays:

Begomovirus Group (NAK 15001) are members of the Geminiviridae family and can infect a wide variety of plants. The viruses are transmitted via insect vectors, whiteflies, and have been known to cause significant crop loss. The nucleic acid hybridization assay for the Begomovirus group was validated to detect the following viruses: Tomato yellow leaf curl virus (TYLCV), Bean dwarf mosaic virus (BDMV), Bean golden mosaic virus (BGMV), Bean golden yellow mosaic virus (BGYMV), Chino del tomate virus (CdTV), Pepper golden mosaic virus (PepGMV), Squash leaf curl virus (SLCV), Tomato golden mosaic virus (TGMV), Tomato mottle virus (ToMoV), and Tomato severe leaf curl virus (ToSLCV).

Curtovirus/Becurtovirus Group (NAK 13100)

Curtovirus/Becurtoviruses are members of the Geminiviridae family and infect a broad range of hosts, including weeds and crops (pepper, tomato, beans, spinach, and cucurbits). Symptoms observed include severe stunting, yellowing, and, if infected early, plants will not produce fruits. Transmitted by leafhoppers and overwintering in weeds, these viruses are becoming a major economic pest. The nucleic acid hybridization assay for the Curtovirus/Becurtovirus group was validated to detect the following viruses: Beet curly top virus (BCTV-A, B, C, E, F, and H), Spinach severe curly top virus (SpSCTAV), and Spinach curly top Arizona virus (SCTAV).
The hybridization assays are offered with AG1 Lysis Buffer and membranes. Processing of the membrane by Agdia Testing Services is included in the cost of the kit. Agdia provides a 1 year warranty and continual customer support.

For more information: info@agdia-emea.com, phone +33 1 60 78 81 64.

Agdia-Emea supports De Ruiter Seed and Seminis Roparun team

Roparun“The Roparun is a relay race of approximately 520 kilometres from Paris and 580 kilometres from Hamburg to Rotterdam, in which people work as a team to give a combined sports performance in order to raise money for people who suffer from cancer. You could also call it an adventure for life. This is also clear from the motto, which has been the same for many years: “Add life to the days, where often days can no longer be added to the life”.
The Roparun of 2015 is the 24th edition. The start takes place in Hamburg and Paris on Saturday the 23rd of May, and both will finish at the same time, on the Coolsingel in Rotterdam on Monday 25th of May.”

Agdia-Emea supports the De Ruiter Seed and Seminis team 162, to follow or support also this team do not hesitate to donate on the following link: http://www.team162.nl/startpagina/

1 2  Scroll to top