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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.
1.1.2. Test samples U.K.
1.1.2.1. Serum samples to be tested, and positive and negative control sera, are diluted 1 in 25 in PBS + 5 % (w/v) skimmed milk + 0,05 % (v/v) Tween 20, 100 μl per well. Incubate for 1 hour at 37 °C. U.K.

For titration, make a twofold dilution series from 1 in 25 (100 μl/well), one serum per plate column, and do the same with positive and negative controls. Incubate for 1 hour at 37 °C.

1.1.2.2. 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.