Validation / Evaluation

In the first part of the QBOL project a survey was set-up to find out wishes and expectations of possible end-users (scientists and technicians of NPPO’s) regarding the data to be generated by QBOL and stored into the Q-bank database.

Objectives

  • To validate developed DNA barcoding protocols and to validate the use of DNA barcoding as a diagnostic tool. 

Tasks

  • Task 10.1  Validation/evaluation of the barcoding protocols
  • Task 10.2  Validation/evaluation of diagnostic use of barcoding
  • Task 10.3  Contingency plan

Deliverables

  • D 10.1 Report of validation of DNA barcoding protocols (Month36)
  • D 10.2 Report of evaluation of diagnostic use of DNA barcoding (Month 36)
  • D 10.3 Scientific publication of results evaluation diagnostic use barcoding (provided that results and time allow this; Month 36)
  • D 10.4 Detailed contingency plan for WP10 (Month 6)

Results

In the first part of the QBOL project a survey was set-up to find out wishes and expectations of possible end-users (scientists and technicians of NPPO’s) regarding the data to be generated by QBOL and stored into the Q-bank database. Based on the end-users’ expectations, and on the queries submitted, the usability of the QBOL database has been improved.

Before the start of the test-performance study in this work package, the developed tests were harmonised as much as possible and a draft EPPO standard ‘DNA barcoding as identification tool for EU regulated plant pests’ has been made. A selection of specimens to be tested in the test-performance study has been made and treated to be non-infectious and non-viable for sending them without permits. A homogeneity test has been performed with all samples before sending. Obtained sequences in the homogeneity test served as standards for comparison with the outcome of the test-performance study.

21 test packages (14 QBOL partners, 7 training sessions) were prepared providing partners and training session organizers with an instruction booklet including the EPPO standard, all DNA purification kits, primers and samples. All results from the QBOL partners were analysed and evaluated in terms of

  1. Number of samples analysed and % of test correct used
  2. % Amplicons obtained.
  3. % Consensus obtained
  4. % Consensus sequence of correct size
  5. % Primers trimmed
  6. Diagnostic sensitivity
  7. Diagnostic specificity
  8. Repeatability
  9. Robustness.

Pitfalls in the use of the EPPO standard, instruction booklet and the use of Q-bank databases have been identified and recommendations for future work have been made.