GENERAL PRINCIPLESU.K.
Optimized protocols for the various methods, validated reagents and details for the preparation of test and control materials are provided in the Appendices. A list of the laboratories that were included in optimization and validation of protocols is provided in Appendix 1.
Since the protocols involve detection of a quarantine organism and will include the use of viable cultures of C. m. subsp. sepedonicus as control materials, it will be necessary to perform the procedures under suitable quarantined conditions with adequate waste disposal facilities and under the conditions of appropriate licences as issued by the official plant quarantine authorities.
Testing parameters must assure consistent and reproducible detection of levels of C. m. subsp. sepedonicus at the set thresholds of the selected methods.
Precise preparation of positive controls is imperative.
Testing according to the required thresholds also implies correct settings, maintenance and calibration of equipment, careful handling and preservation of reagents and all measures to prevent contamination between samples, e.g. separation of positive controls from test samples. Quality control standards must be applied to avoid administrative and other errors, especially concerning labelling and documentation.
A suspected occurrence, as referred to in article 4(2) in directive 93/85/EEC implies a positive result in diagnostic or screening tests performed on a sample as specified in flow charts.
If the first screening test (IF or PCR/FISH) is positive, then contamination with Cms is suspected and a second screening test must be done. If the second screening test is positive, then the suspicion is confirmed (suspected occurrence) and the testing according to the scheme must be continued. If the second screening test is negative, then the sample is considered not contaminated with Cms.
Therefore a positive IF test as referred to in article 4(2) is defined by a positive IF reading confirmed by a second screening test (PCR/FISH).
Confirmed presence as referred to in article 5(1) in directive 93/85/EEC implies the isolation and identification of a pure culture of C. m. subsp. sepedonicus with confirmation of pathogenicity.