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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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Changes over time for: Division 2.3.

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EU Directives are published on this site to aid cross referencing from UK legislation. Since IP completion day (31 December 2020 11.00 p.m.) no amendments have been applied to this version.

[F1 [F2 [F32.3. Complement fixation test (CFT) U.K.

2.3.1. The antigen represents a bacterial suspension in phenol-saline (NaCl 0,85 % (m/v) and phenol at 0,5 % (v/v)) or in a veronal buffer. Antigens may be delivered in the concentrated state provided the dilution factor to be used is indicated on the bottle label. The antigen must be stored at 4 °C and not frozen. U.K.
2.3.2. Serums must be inactivated as follows: U.K.
  • bovine serum: 56 to 60 °C for 30 to 50 minutes,

  • porcine serum: 60 °C for 30 to 50 minutes.

2.3.3. In order to carry out the genuine reaction within the test procedure, a complement dose higher than the minimum necessary for total haemolysis shall be used. U.K.
2.3.4. In carrying out the complement fixation test, the following controls must be made each time: U.K.
(a)

control of the anti-complementary effect of the serum;

(b)

control of the antigen;

(c)

control of sensitised red blood cells;

(d)

control of the complement;

(e)

control using a positive serum of sensitivity at the start of the reaction;

(f)

control of the specificity of the reaction using a negative serum.

2.3.5. Calculation of results U.K.

The OIEISS contains 1 000 international CFT units (ICFTU) per ml. If the OIEISS is tested in a given method the result is given as a titre (i.e. highest direct dilution of the OIEISS giving 50 % haemolysis, T OIEISS ). The test result for the test serum given as titre (T TESTSERUM ) must be expressed in ICFTU per ml. In order to convert the expression of a titre into ICFTU, the factor (F) necessary to convert a titre of an unknown test serum (T TESTSERUM ) tested by that method into the ICFTU expression can be found from the formula:

F = 1 000 × 1/T OIEISS

and the content of international CFT units per ml of test serum (ICFTU TESTSERUM ) from the formula:

ICFTU TESTSERUM = F × T TESTSERUM

2.3.6. Interpretation of results U.K.

A serum containing 20 or more ICFTU per ml shall be considered to be positive.] ] ]

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