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[F1ANNEX IV U.K. AFRICAN HORSE SICKNESS DIAGNOSIS

PART A U.K. Serological tests

1. Indirect ELISA for the detection of antibodies to African horse sickness virus (AHSV) U.K.

1.1. Test procedure U.K.
1.1.1. Solid phase U.K.
1.1.1.1. Coat ELISA plates with recombinant AHSV-4 VP7 diluted in carbonate/bicarbonate buffer, pH 9,6. Incubate plates overnight at 4 °C. U.K.
1.1.1.2. Wash the plates five times with distilled water containing 0,01 % (v/v) Tween 20 (washing solution). Gently tap the plates onto absorbent material to remove any residual wash. U.K.
1.1.1.3. Block the plates with phosphate buffered saline (PBS) pH 7,2 + 5 % (w/v) skimmed milk (Nestlé Dry Skim Milk TM ), 200 μl/well, for 1 hour at 37 °C. U.K.
1.1.1.4. Remove the blocking solution and gently tap the plates onto absorbent material.] U.K.