F1ANNEX ITEST SCHEME FOR DIAGNOSIS, DETECTION AND IDENTIFICATION OF THE RING ROT BACTERIUM, CLAVIBACTER MICHIGANENSIS (Smith) Davis et al. ssp. SEPEDONICUS (Spieckermann et Kotthoff) Davis et al.

SCOPE OF THE TEST SCHEME

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Amendments (Textual)
Appendix 4Determination of contamination level in IF and FISH tests
  1. 1.

    Count the mean number of typical fluorescent cells per field of view (c)

  2. 2.

    Calculate the number of typical fluorescent cells per microscope slide window (C)

    C = c × S/s

    where

    S

    =

    surface area of window of multispot slide, and

    s

    =

    surface area of objective field.

    s = πi2/4G2K2

    where

    i

    =

    field coefficient (varies from 8 to 24 depending upon ocular type)

    K

    =

    tube coefficient (1 or 1,25)

    G

    =

    magnification of objective (100x, 40x etc.).

  3. 3.

    Calculate the number of typical fluorescent cells per ml of resuspended pellet (N)

    N = C × 1 000/y × F

    where

    y

    =

    volume of resuspended pellet on each window, and

    F

    =

    dilution factor of resuspended pellet.