Council Directive of 26 June 1964 on animal health problems affecting intra-Community trade in bovine animals and swine (64/432/EEC)

[F1 C. Enzyme-linked immunosorbent assay (Elisa) for detecting enzootic bovine leukosis U.K.

1. The material and reagents to be used are as follows: U.K.

(a)

solid-phase microplates, cuvettes or any other solid phase;

(b)

the antigen is fixed to the solid phase with or without the aid of polyclonal or monoclonal catching antibodies. If antigen is coated directly to the solid phase, all test samples giving positive reactions have to be retested against control antigen in the case of EBL. The control antigen should be identical to the antigen except for the BLV antigens. If catching antibodies are coated to the solid phase, the antibodies must not react to antigens other than BLV antigens;

(c)

the biological fluid to be tested;

(d)

a corresponding positive and negative control;

(e)

conjugate;

(f)

a substrate adapted to the enzyme used;

(g)

a stopping solution, if necessary;

(h)

solutions for the dilution of the test samples for preparations of the reagents and for washing;

(i)

a reading system appropriate to the substrate used.

2. Standardization and sensitivity of test U.K.

The sensitivity of the Elisa assay must be of such a level that E4 serum is scored positive when diluted 10 times (serum samples) or 250 times (milk samples) more than the dilution obtained of individual samples when these are included in pools. In assays where samples (serum and milk) are tested individually E4 serum diluted 1 to 10 (in negative serum) or 1 to 250 (in negative milk) must be scored positive when tested in the same assay dilution as used for the individual test samples. The official institutes listed in A.2 will be responsible for checking the quality of the Elisa method, and in particular for determining, for each production batch, the number of samples to be pooled on the basis of the count obtained for the E4 serum.

The E4 serum will be supplied by the National Veterinary Laboratory, Copenhagen.

3. Conditions for use of the Elisa test for EBL U.K.

The Elisa method may be used on a sample of milk or whey taken from the milk collected from a farm with at least 30 % of dairy cows in milk.

If this method is used, measures must be taken to ensure that the samples taken can be identified with the animals from which the milk or sera examined were taken.]

Textual Amendments

F1 Substituted by Council Directive 97/12/EC of 17 March 1997 amending and updating Directive 64/432/EEC on health problems affecting intra-Community trade in bovine animals and swine.