Work Package 1

WP1 focus on validation of tests using different methods (i.e. (real-time) PCR, LAMP, ELISA, LFD) according to EPPO Standard PM 7/098, but improved especially by including a statistical approach developed in the frame work of WP2.

Performance criteria of several tests will be determined by organizing interlaboratory test performance studies (TPS) according to EPPO Standard PM 7/122 in which participating laboratories will use the same test for analysing identical sets of reference samples simulating the samples routinely analysed.

Two rounds of tests performance studies will be organised.

• The first round will be organized in early 2019 for six prioritized pests (Erwinia amylovora, Pantoea stewartii, citrus tristeza virus, plum pox virus, Fusarium circinatum and Bursaphelenchus xylophilus) in a range of matrices and for a range of methods.
• The second round will be performed in 2020 on pests and associated tests based on the needs of stakeholders and to the market analysed in the framework of WP4.

Erwinia amylovora

List of deliverables for WP1

• 1.1. Report detailing the minimum performance parameters to select tests for validation and selection of laboratories for TPS
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1.1. Report detailing the minimum performance parameters to select tests for validation and selection of laboratories for TPS

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For each pest, criteria for selection of tests for the TPS have been set. These criteria have been divided in five groups:

1. validation data (e.g. availability of data on analytical sensitivity and specificity, repeatability, reproducibility…),
2. applicability (e.g. can the test be used with different matrix?, are ready to use reagents and controls available?…),
3. protocols (e.g. are they available protocols? How long does the test take to perform the test?...),
4. chemicals (e.g. availability, stability…) and
5. equipment (e.g. are specific equipments needed? cost of those equipments?…).




Criteria for selection of participants for the TPS have also been set. Amongst the most important criteria for selection for participants of TPS are technical expertise for the pest group and the method, authorization to work with the specific pest and that the participating laboratory has quality assurance in place.
These criteria enable evaluation of whether participants are proficient to perform the tests, have the necessary equipment and a permit to work with regulated organism.

For the purpose of setting the weight for each criterion for a specific pest, the scope of the testing (e.g. use for detection or screening, symptomatic or asymptomatic material, selection of methods…) for specific pests was set and common rules for each selection process were defined.

For the 6 priority pests for the first TPS, 41 tests in total and 17 to 32 laboratories per pests were selected (see details on the TPS on TPS page)

Partners involved : NIB, NVWA, FERA, ANSES

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• 1.2. List of tests for validation - Round 1
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1.2. List of tests for validation - Round 1 ×

For each of the 6 pests of the first round of TPS, a list of methods and tests was selected for validation. This selection was based on the scope and criteria prepared in D1.1.

Pest	Scope of the TPS	Methods and tests selected
Erwinia amylovora	detection in symptomatic plant material	2 real-time PCR, 1 LAMP, 2 LFD
Pantoea stewartii subsp. stewartii	detection in asymptomatic plant material (maize seeds)	2 PCR, 4 real-time PCR
Citrus tristeza virus	detection in symptomatic plant material and in plant material spiked with the virus in order to mimic asymptomatic samples	1 RT-PCR, 2 real-time RT-PCR, 5 ELISA, 1 Immunostrip, 1 LAMP, 1 TPIA
Plum pox virus	detection in symptomatic and asymptomatic leaf material of Prunus spp.	2 RT-PCR, 3 real-time RT-PCR, 3 DAS-ELISA
Fusarium circinatum	identification from culture material using plating methods and detection from culture material and DNA extracts using molecular methods	1 plating, 1 PCR, 4 real-time PCR
Bursaphelenchus xylophilus	detection in asymptomatic plant material and its identification	2 PCR, 2 real-time PCR, 1 LAMP


Partners involved : NIB, NVWA, FERA, ANSES

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• 1.3. List of tests for validation - Round 2
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1.2. List of tests for validation - Round 2 ×

A list of methods and tests for validation for test performance study (TPS) Round 2 (for laboratory and/or on-site use) was prepared for six selected pests: Xanthomonas citri pv. citri, Xylophilus ampelinus, Cryphonectria parasitica, tomato spotted wilt tospovirus, tomato brown rugose fruit virus and plum pox virus. The listed tests were first validated in preliminary studies by TPS organizers in order to select the final tests for TPS, based on the scope and criteria also described in this deliverable.



Pest	Scope of the TPS	Tests included in preliminary studies	Tests selected for TPS (methods)
Xanthomonas citri pv. citri	detection of X. citri pv. citri in leaves and fruits	20	13 (PCR, real-time PCR, LAMP-PCR, direct molecular tests performed from ImmunoStrips® (Agdia) and WhatmanTM FTA cards)
Xylophilus ampelinus	detection and identification of X. ampelinus in grape vine material	10	9 (IF, ELISA, PCR, real-time PCR)
Cryphonectria parasitica	detection of C. parasitica in wood material	3	3 (PCR, real-time PCR)
Tomato spotted wilt virus	detection and identification of TSWV in symptomatic leaves of tomato	19	8 (ELISA, LFD, RT-PCR, real-time RT-PCR)
Tomato brown rugose fruit virus	detection of ToBRFV in leaves and fruits of tomato and pepper	12	5 (RT-PCR, real time RT-PCR)
Plum pox virus (on site testings)	detection of PPV in leaves	4	3 (LFD RPA, LFD)


Partners involved : ANSES, CREA, FERA, NIB, UNITO

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• 1.4. TPS reports with description of the method, materials and software used, as well as the data analysis - Round 1
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• 1.5. TPS reports with description of the method, materials and software used, as well as the data analysis - Round 2
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Once the deliverables will be finalised, they will be available here for download.

Work Package 2

This WP aims to improve validation approaches for diagnostic technologies to maximise their usefulness for users (diagnosticians) and decision-makers (at National, European or Regional levels) and their use in routine diagnostics. In details, the WP aims at:

1. Improving the current EPPO Standards for validation of tests for plant pest diagnostics (PM 7/098) and for the performance on interlaboratory comparisons (PM 7/122) by incorporating new statistical tools and predictive model;
2. Developing best practice guidelines;
3. Improving generic approaches for the validation and developing best practice guidelines for the validation and application of non-targeted (generic) diagnostic procedures, using high throughput sequencing (HTS or NGS) technologies for viruses detection, in plant pest diagnostics as a model.

List of deliverables for WP2

• 2.1. Guidelines for the revision of the EPPO Standards PM 7/098 and PM 7/122 for validation studies (including TPS)
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2.1. Guidelines for the revision of the EPPO Standards PM 7/098 and PM 7/122 for validation studies (including TPS) ×

One of the objectives of VALITEST is to propose an improvement of the validation approach for diagnostic tests. To that effect, WP2 has reviewed statistical methods and has written a report presenting a new possible approach using wherever possible and whenever applicable, statistical methods recommended by international standards with examples of their application to plant health diagnostic tests.

For the most important test performance criteria, the following statistical methods have been identified.

Performance criteria/Parameter	Statistic method for the calculation of the parameters
Analytical sensitivity	Probability of detection
Diagnostic sensitivity	As per EPPO PM 7/98 (4)
Diagnostic specificity	As per EPPO PM 7/98 (4)
Repeatability	Accordance
Reproducibility	Concordance









Figure 1: Using generalized linear model to calculate the probability of detection for different methods based on the dilution level

For a probability of detection of 75%,
Method 6 is better while for
a probability of detection of 95%,
Method 4 is better.




Those statistical methods will be evaluated during the VALITEST project and also by EPPO diagnostic Panels and proposed for possible inclusion in the EPPO PM 7/98. It is also proposed to create an online application for easy analysis of the data collected during intra and inter-laboratory studies.

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• 2.2. “Best practice” guidelines for validation and routine use of non-targeted techniques in diagnostic setting which could serve as a basis for a new EPPO Standard
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• 2.3. Scientific publication of the results of the validation of approach applied on 5 existing datasets
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Work Package 3

WP3 aims at developing recommendations for the quality of reference materials for validation purposes.

By building on existing guidelines provided by Q-collect and EPPO as well on the databases and collections present among the partners of the project’s consortium, WP3 will establish and disseminate (through WP6 and WP7) quality guidelines for the production and dissemination of reference materials for use in phytosanitary tests including possible quantification of targets in reference material for validation purpose. This will help in improving the overall standard of tests for plant pests in Europe and allow for a further implementation of shared standards and will introduce metrology principles in plant pest diagnostics.

Different types/classes of reference material (i.e. quantitative vs. qualitative) each have their own production characteristics thus require different guidelines. In consultancy with the end-users and building on the information already collected with Q-collect, these different needs and requirements will be defined and existing methods suitable for their characterization, including their commutability and the extent to which they need to resemble samples, will be evaluated and if necessary refined. The recommendations developed in this WP3 will be evaluated during the production of reference material in WP1.

In this way the guidelines will ensure that the reference materials produced shall match (as closely as possible) the plant materials encountered in routine testing both in terms of plant matrices as well as with respect to the infestation level of the specific pest.

List of deliverables for WP3

• 3.1. List of the criteria the reference materials have to meet for use in validation studies
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3.1. List of the criteria the reference materials have to meet for use in validation studies ×



A list of general minimum criteria for the production of reference materials (RMs) to be used in interlaboratory studies (including validations through test performance studies (TPS)) was developed. The list comprises criteria previously identified in international standards (ISO) and guidelines (EPPO), in deliverables of relevant projects and in other sources as well as additional criteria based on partners own experience.





The identified criteria are the intended use of the material (for example TPS for detection methods for Erwinia amylovora in symptomatic plant material), its identity, traceability, commutability level (i.e. similarity to the actual sample), homogeneity, stability, assigned value (i.e. expected result of the test) and purity. Where relevant, each criterion was first defined as a series of levels from the highest to lowest ranking with the lowest ranking considered to be the minimum. Depending on the intended use, the RMs may need to fulfil higher levels of selected criteria or some criteria may not be relevant at all.


The criteria identified in this work are applicable, potentially with minor modification, beyond the scope of this project to other pests and other uses of RMs.

Partners involved : WR, UNITO, WBF, EPPO, NIB, NVWA, ANSES, IPADLAB, SEDIAG

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• 3.2. Development draft Standard Operating Procedures (SOPs) available for the production of the reference materials identified in Task 3.1
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3.2. Development draft Standard Operating Procedures (SOPs) available for the production of the reference materials identified in Task 3.1 ×



Graphical presentation of the SOP










A general standard operating procedure (SOP) for the production of reference material (RM) for use in plant health diagnostics was developed. The general SOP was designed based on (limited) information on existing SOPs and guidelines available with the consortium partners.

The general SOP describes the different steps required in the production process, ranging from the different possible sources of the reference material (e.g. field material, working collection, reference material or certified reference material), tests to confirm its identity, possibly required multiplication steps to the actual production process. For each step in the process, criteria and critical points are identified. The criteria that reference material has to meet, and their minimum required levels as identified and described in Deliverable 3.1 of WP3 form an import basis of the final production process.

Partners involved : WR, UNITO, WBF, EPPO, NIB, NVWA, ANSES, IPADLAB, SEDIAG

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• 3.3. Guidelines and Standard Operating Procedures (SOP) finalised for the production of the reference materials
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Work Package 4

Work package 4 will assess the demand for current and future tests and operating procedures. Stakeholder feedback will be directly incorporated into the project through WP4, as it will form the basis for the prioritisation of tests in round two of the Test Performance Studies.

Through incorporating elements of the multi-actor approach and co-design with end-users, this approach will support plant health policies by ascertaining stakeholder views on attributes that lead to the adoption of surveillance and testing tools.
Furthermore, several market impact assessments will be conducted in close collaboration with WP7, to assess the scale of the effect of selected tests or toolkits in terms of monetised costs and benefits. A cost benefit approach (CBA) will be adopted, which will identify who benefits, when and at what scale; this information can then be used to assess the market potential for tests and toolkits.

List of deliverables for WP4

• 4.1. Report on stakeholder priorities for tests and general prioritization framework
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4.1. Report on stakeholder priorities for tests and general prioritization framework ×

Main aim of work-package 4 is a better understanding of the demands for current and future testing options. This is to be achieved via a qualitative assessment of stakeholder requirements using online surveys supplemented by desk-based research, as well as impact assessments. To make the most of work-package 4 and to ensure that Test Performance Study (TPS) targets are determined considering the latest developments in demand, it was decided - prior to the project start - to use the results from the stakeholder survey for prioritising target pests. This also reflects the importance of co-design and the multi-actor approach, which are central to this project.

Deliverable 4.1 reports on identified stakeholder priorities for tests as well as the general prioritisation framework, which was created in collaboration with all affected work-packages. After applying the framework to the long list of target pests obtained through the surveys, the selection of pests for TPS round 2 was the following: Arabis mosaic virus, Cryphonectria parasitica, Melampsora medusa, Plum pox virus, Tobacco ringspot virus, Tomato brown rugose fruit virus, Tomato ringspot virus, Tomato spotted wilt virus, Xanthomonas citri pv citri, Xylophilus ampelinus. Of those the following have been selected for the second round of TPS: Cryphonectria parasitica (UNITO), Tomato brown rugose fruit virus (CREA), Plum pox virus (ANSES), Tomato spotted wilt virus (NIB), Xanthomonas citri pv citri (ANSES), Xylophilus ampelinus (FERA).





Partners involved : all partners

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• 4.2. Report on economic impact of priority tests
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Work Package 5

To ensure the proper use of a validated test in time, a monitoring of its use is required. The current approach is based on pest specific proficiency tests, it requires each laboratory to show its proficiency for each pest and test. Considering the difficulties of organising proficiency tests (time, cost, material availability, and complexity), all the pests and tests cannot be covered. This approach is no longer adequate and sustainable.

In VALITEST, experts will study the possibility to evaluate the proficiency of laboratories on tests (which have sufficient commonality in process and procedure) through a single PT. The needs of laboratories for PT will be collected and analysed in order to prepare guidelines on “horizontal” PT.

This action will provide guidelines which will be available for discussions with accreditation bodies.

List of deliverables for WP5

• 5.1. Analysis of the needs of the laboratories and applicability of the horizontal proficiency testing approach based on the questionnaire answers
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5.1. Analysis of the needs of the laboratories and applicability of the horizontal proficiency testing approach based on the questionnaire answers ×






The 5th work package of the VALITEST project aims at developing guidelines on a horizontal approach allowing the laboratories to undertake proficiency testing (PT) without having to participate in proficiency tests for all the tests they use.


A range of data has been collected using three different approaches, i) the study of accreditation scopes of some laboratories involved in diagnostics, ii) a workshop with diagnostic experts, iii) a survey sent to laboratories listed in the EPPO database on diagnostic expertise. The data collected allow a better understanding of the expectations of laboratories, of what they would consider as acceptable and of the applicability of the horizontal proficiency testing approach.

According to the data collected, a PT plan should be conducted over 3 to 4 years. Each year a single PT should be organized per field. It could contain several pests, however, to limit the workload no more than 3 tests should have to be performed on a maximum of 10 samples. The PT should include whenever possible the different pests included in the scope of accreditation of the laboratories, but also emerging pests. The pests to consider as a priority have been identified. PT samples should be as similar as possible to routine samples, ideally different matrices should be included in each PT. Among the different factors determining the reliability of a test, the design of the PT plan should especially focus on human factors (i.e. the proficiency of the staff to use the different techniques). The laboratories do not wish to spend more than 3 500 € per year for a service providing proficiency tests covering all their needs.



Partners involved: ANSES, WBF, EPPO, FERA, NIB, ULG, NVWA, CREA

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• 5.2. Guidelines on an approach to undertake horizontal proficiency testing
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Work Package 6

WP6 will be focused on communication, training and dissemination of the validation data and activities performed within the project. It will target the different identified stakeholder groups (including researchers, policy makers, inspectors, advisory services, diagnostic laboratories, industries) in order to facilitate the take-up and successful exploitation of the project results.

Training workshops on the guidance and frameworks developed in VALITEST will be carried out for diagnosticians in order to harmonise validation processes throughout the region and to ensure that they clearly understand and benefit from the project outputs. These Workshops will be organized in collaboration with WP7. Dissemination and training workshops will also be completed by the development of webinars and video tutorials on the validation process. Training for inspectors will also be included in the programme of EPPO Workshops for inspectors, and contact will be made with the EU Commission and coordinators of the programme Better Training for Safer Food (BTSF) to encourage synergies.

New validation data generated, or existing validation data gathered during the project will be made available through an improved version of the EPPO Database on diagnostic expertise (section validation data http://dc.eppo.int/validationlist.php ).

Finally, dissemination through scientific publications, technical articles and participations in appropriate events and congresses, will be prepared to present the different outcomes of the project.

EPPO Diagnostic Expertise Database

List of deliverables for WP6

• 6.1. Project website and social media accounts
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• 6.2. Survey on the needs of users of the EPPO Database and exploitation of the data
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6.2. Survey on the needs of users of the EPPO Database and exploitation of the data ×

One of the objectives of VALITEST is to improve the searching capacity of the validation section of the EPPO Database on Diagnostic expertise to ensure optimal use. In order to evaluate the needs of users a survey was elaborated and sent to diagnostic experts and industry contacts.






179 responses were received corresponding to an answer rate of 36%.



Map of responding countries
(one response from the United States was also received)

Respondents pointed out different queries that they would like to be able to make, such as querying the database based on the plant species, the type of samples, the objective of the tests (detection/identification)… It appears that multiple queries (for example : pest + method + plant species) are of interest for most respondents. Besides, most respondents are in favor of subdividing methods, in particular serological and molecular method, into more categories.

These results will be used to identify and plan future IT developments necessary to establish the new features of the validation section. The new version of the EPPO Database on diagnostic expertise will be launched at the end of the VALITEST project.

Partners involved : all partners + external consultation via the EPPO network.

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• 6.3. Inventory of validation data available
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6.3. Inventory of validation data available ×

A survey was conducted with the objective to gather information on the validation data available for tests of the first test performance study (WP1).





43 different laboratories (65% of the laboratories contacted) provided validation data.



Map of responding countries

297 descriptions of tests were provided by laboratories for the 6 pests as follows: Erwinia amylovora (102), Pantoea stewartii subsp. stewartii (42), Citrus tristeza virus (29), Plum pox virus (47), Fusarium circinatum (34) and Bursaphelenchus xylophilus (43). 70 were described as validated, out of which 20 were already uploaded in the EPPO Database on Diagnostic expertise. 10 new validation reports were collected and will be entered in the same database.

Partners involved : all partners + external consultation via the EPPO network.

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• 6.4. Dissemination and Training plan
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6.4. Dissemination and Training plan ×

The training and dissemination plan presents the objectives of training and disseminations and the tools provided to ensure that the objectives will be met.

The objectives of VALITEST dissemination and training activities can be summarised as follows:

• Contributing to the wider harmonization of the validation process
• Sharing validation data generated during the project
• Raising awareness and capacity building among stakeholders
• Enhancing product development by the diagnostics industry
• Increasing the project visibility
• Contributing to promote EU research




Different stakeholders have been identified as well as the key message to deliver and the dissemination tools to be developped to that end.

Target groups	Key messages	Main Dissemination tools
Consumers and Society (general public)	Project objectives and results, with a particular focus on pest impacts and benefits of validation data to plant health	Project website, brochure, leaflets, social media accounts, general press articles, etc.
Farmers, landowners, agricultural advisors, breeders	Project objectives and results; practical solutions for pest detection (e.g. on-site tools) to support pest management; training on tests.	Project website, brochure, leaflets, social media accounts, general press articles, training toolkit, workshops
Inspection services, NPPOs, Policy and decision makers	Project objectives and results; practical solutions for pest detection to support pest management; training on tests. Guidelines (validation, proficiency evaluation)	Project website, brochure, leaflets, social media accounts, technical and scientific articles, training toolkit, invitation to workshops
Scientific community	Validation data, Knowledge of end-users’ needs (Innovation-driven research)	Scientific papers, posters, conferences and workshops, training toolkit, targeted communication, invitation to project meetings and workshops
Industries	Project objectives and results; new tools and procedures for diagnostics validation; evaluation of consumer perception; acceptability of novel products; business plan to support product development	Project website, brochure, leaflets, technical articles, social media accounts, direct communication by project partners




Partners involved: all partners

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• 6.5. Information materials (project fact-sheet, flyers, poster)
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6.5. Information materials (project fact-sheet, flyers, poster) ×

Project fact-sheets, flyers and poster are available for download in the publication section of the website : https://www.valitest.eu/publications




Partners involved: all partners

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• 6.6. Report on improved version of the validation section of the EPPO Database on diagnostic expertise, including new data provided by laboratories
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6.6. Report on improved version of the validation section of the EPPO Database on diagnostic expertise, including new data provided by laboratories ×

The EPPO Database on Diagnostic Expertise has been transferred to a new redesigned and more user-friendly database on the 4th of May 2020.

Access the EPPO Database on Diagnostic Expertise – https://dc.eppo.int

The EPPO Database on Diagnostic expertise includes a specific section on validation data for diagnostic tests. Laboratories can deposit online validation data that they have generated on specific tests and these can be made visible to all users of the database. The validation data can then be retrieved from the database in the form of a PDF file including the description of the test evaluated (pest x matrix x method) and the performance data related to the test (the information is presented in a harmonized format).

One of the objectives of VALITEST was to improve the searching capacity of the database to ensure optimal use. In order to evaluate the needs of users a survey was organized (see deliverable 6.2). Further needs were identified during meetings with users (e.g. EPPO Panels on diagnostics, mid-term conference). The features identified have been implemented in the database and include:

• Allow searches to be made using key words for specific descriptors such as pest, method, plant species, matrix, EPPO-IPPC test
• Allow combined and flexible queries to be made and, in particular, multiple pest queries
• Allow sorting of information within different methods, in particular for molecular and serological methods
• Allow searches of tests used for detection, identification or both
• Allow searches to be made on kits with the possibility to specify whether the kit was used according to manufacturer’s instructions
• Provide examples on the way EPPO would like laboratories to enter their data.


This transfer allowed the improvements of all the sections of the website. Although the focus was on the section on validation data, data on laboratories and expertise are now also more easily searchable.

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• 6.8. Report on the organisation of training activities (webinars, video tutorial, practical training sessions) (merge of original D6.7. Workshop for laboratories and D6.8. Webinar and video tutorials on the validation process)
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• 6.9. Final Plan for the Dissemination and Exploitation of project Results (PDER)
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Once the deliverables will be finalised, they will be available here for download.

Work Package 7

WP7 will be dedicated to the promotion of the project’s results and market exploitation.

This WP, including all SME of the consortium, will promote the results of the project in connection with their commercial offer.

It will prepare the foundations for an EU Association of the Plant Health Diagnostics Industry and an EU Plant Health Diagnostics Charter for Industry. The drafting of these elements will be done while verifying that their application will be coherent with the SMEs’ business constraints.

Additionally, WP7 will contribute to dissemination workshops presenting use of kits validated in the framework of this project.

In association with WP4, WP7 will identify from the stakeholders needs, and the results of the projects for diagnostic tests that are not commercially available, the potential for developing new kits validated to consensus standards that could be swiftly brought to market. These new kits will be useful in field monitoring, eradication and containment campaigns for producers or competent authorities, and help to minimize economic and environmental damage as further explained in Section 2.

List of deliverables for WP7

• 7.1. Results exploitation plan
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7.1. Results exploitation plan ×

WP7 is dedicated to the promotion of the project’s results and market exploitation.

Exploitation refers to the use of the results of the project. This definition does not have a pure commercial meaning, thus opening the scope of application of results at different levels and in different domains. This leads to both commercial and non-commercial exploitation. While commercial exploitation is more related to taking results to the market, non-commercial exploitation is more related to the effective use of knowledge, know-how, methodologies or standards.




Dissemination and exploitation goals




The exploitation plan aims at defining an effective long-term strategy to pursue the following objectives:

• Explore and assess application areas to facilitate the exploitation of the project’s results;
• Set the foundations for further opportunities to ensure the achievement of impact during and after the end of the project.


The key exploitable results and exploitation routes have been identified for several work packages (WP1, WP5 and WP6). For those work packages, the results of the project could be used for further research activities, for creating and providing services and in standardisation activities.

Besides two general exploitation guidelines (one for industrial partners and one for research institution partners) have been developed to help partners reach the expected exploitation of the results.

Partners involved : IPADLAB, WBF, BIOREBA, LOEWE, EPPO, NIB, ULG, CD, WR (Prime Diagnostics), SEDIAG, GIORIN

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• 7.2. Communication and Exploitation strategy plans

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• 7.3. Definition of the EU Plant Health Diagnostics Charter for Industry
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• 7.4. Report on education and training session during the project
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Once the deliverables will be finalised, they will be available here for download.

Work Package 8

The objective of the project’s management is to guarantee the correct implementation of the activities, the achievement of the project objectives and expected results.