For a holding to qualify for brucellosis-free status, testing for brucellosis (B. melitensis) is performed by means of the Rose Bengal method or by the complement-fixation method described in the Annex to Decision 90/242/EEC or by any other method recognized in accordance with the procedure laid down in Article 15 of this Directive. The complement-fixation method is used for tests on individual animals.

When carrying out the Rose Bengal test, if more than 5 % of the animals on a holding show a positive reaction, a further test is carried out on every animal on the holding by means of the complement-fixation method.

Serum containing 20 or more ICFT units/ml must be regarded as positive in the complement-fixation test.

The antigens used must be approved by the national laboratory and must be standardized against the second international standard anti-brucella abortus serum.