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Council Directive of 26 June 1964 on animal health problems affecting intra-Community trade in bovine animals and swine (64/432/EEC)
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- Directives originating from the EU
- 1964 No. 432
- ANNEX C
- Division 2.
- Division 2.6.
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Changes over time for: Division 2.6.
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[F1 [F2 [F32.6. Serum agglutination test (SAT) U.K.
2.6.1. The antigen represents a bacterial suspension in phenol-saline (NaCl 0,85 % (m/v) and phenol at 0,5 % (v/v)). U.K.
Formaldehyde must not be used.
Antigens may be delivered in the concentrated state provided the dilution factor to be used is indicated on the bottle label.
EDTA may be added to the antigen suspension to 5 mM final test dilution to reduce the level of false positives to the serum agglutination test. Subsequently the pH of 7.2 must be readjusted in the antigen suspension.
2.6.2. The OIEISS contains 1 000 international units of agglutination. U.K.
2.6.3. The antigen shall be prepared without reference to the cell concentration, but its sensitivity must be standardised in relation to the OIEISS in such a way that the antigen produces either a 50 % agglutination with a final serum dilution of 1/600 to 1/ 1 000 or 75 % agglutination with a final serum dilution of 1/500 to 1/750. U.K.
It may also be advisable to compare the reactivity of new and previously standardised batches of antigen using a panel of defined sera.
2.6.4. The test shall be performed either in tubes or in microplates. The mixture of antigen and serum dilutions shall be incubated for 16- to 24-hours at 37 °C. U.K.
At least three dilutions must be prepared for each serum. Dilutions of suspect serum must be made in such a way that reading of the reaction at the positivity limit is made in the median tube (or well for the microplate method).
2.6.5. Interpretation of results: U.K.
The degree of Brucella agglutination in a serum must be expressed in IU per ml.
A serum containing 30 or more IU per ml is considered to be positive.] ] ]
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.
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