TABLE CONTENTS

Quality Requirements for Accreditation and Standard Operating
Procedures: What could be a Reference Method for Quarantine Nematodes
Detection?
Anthoine, G. (1), L. Ladeveze (1), V. Gaar (2), N. Viane (3) & S. Hockland (4)
Laboratoire National de la Protection des Végétaux, Unité de nematologie, Domaine de la Motte, BP 35327,
35653 LE RHEU cedex, France; (2) State Phytosanitary Administration, Diagnostic Laboratory, Drnovska
73/507, 161 06 PRAHA 6, Czech Republic ; (3) Institue for Agricultural and Fisheries Research, Crop
Protection Unit, Burg. Van Gansberghelaan 96-bus2, 9820 Merelbeke, Belgium; (4) Central Science Laboratory,
Pest and Disease Identification Team, Plant Health Group, Sand Hutton, York Y041 1LZ, United Kingdom.
The recent increase of trade in plants has stressed the need for standard operating procedures
to get reliable and comparable results among diagnostic laboratories, preventing major
dispute. For quarantine plant-parasitic nematodes, regional or international diagnostic
protocols are already produced: e.g. IPPC international protocols or EPPO regional protocols.
Even if helpful, these protocols often come closer to a compilation of most known or used
tests, without elements about their performance (sensitivity, specificity, repeatability,
reproducibility). These validation elements become more and more demanded as diagnostic
laboratories develop quality management system and need to prove the validity of the method
in use (accreditation - ISO 17025 standard). Thus consistent discussions began with different
diagnostic protocols. Examples are given here from the IPPC protocol for Bursaphelenchus
xylophilus with the assessment of the molecular identification. The specificity of published
tests (PCR, PCR-RFLP) was evaluated against a set of B. xylophilus, B. mucronatus and other
Bursaphelenchus species, chosen to represent part of the genetic diversity described. In
practice most of the tests are reliable when compared to published results. Another example
is given from the EPPO protocol for Xiphinema americanum and the limits of the different
detection or identification methods. Performance of the different extraction methods would
be discussed. Limits of the morphological or molecular identifications and the reliability of
the final results would be presented and analysed. These results highlight that a diagnostic
protocol should inform about the performance of the tests included to avoid mistakes and the
best route to ensure reliable results (need for complementary tests). Moreover, at a regional or
international level, definition of minimum performance criteria should be discussed and a
validation process might be described. Then diagnostic protocols would offer more reliability
and ease for all diagnostic laboratories undertaking official analysis, whether they are
accredited or not.
5 th International Congress of Nematology, 2008 74