Commission Delegated Regulation (EU) 2020/689Show full title

ANNEX III U.K. DIAGNOSTIC METHODS FOR THE GRANTING AND MAINTENANCE OF DISEASE-FREE STATUS FOR CERTAIN DISEASES OF TERRESTRIAL ANIMALS

Section 1 U.K. Infection with Brucella abortus, B. melitensis and B. suis

1. Serological tests U.K.

(a)

tests for blood samples

(i)

buffered Brucella antigen tests;

(ii)

complement fixation test (CFT)

(iii)

indirect enzyme-linked immunosorbent assay (I-ELISA)

(iv)

fluorescence polarisation assay (FPA)

(v)

competitive enzyme-linked immunosorbent assay (C-ELISA)

(b)

tests for milk samples

(i)

ring test (MRT)

(ii)

I-ELISA

2. Brucellin skin test (BST) U.K.

For the testing as referred to in section 1 and 2 of Chapter 1 of Part I of Annex IV, Brucellin skin test (BST) shall only be used in ovine and caprine animals.

Section 2 U.K. Infection with mycobacterium tuberculosis complex

1. Tuberculin skin tests U.K.

(a)

the single intradermal tuberculin test (SITT)

(b)

the comparative intradermal tuberculin test (CITT)

2. Gamma-interferon assay U.K.

Section 3 U.K. Enzootic bovine leukosis

1. Serological tests U.K.

(a)

tests for blood samples

(i)

agar gel immuno-diffusion test (AGID)

(ii)

blocking enzyme-linked immunosorbent assay (B-ELISA)

(iii)

I-ELISA

(b)

tests for milk samples

(i)

I-ELISA

Section 4 U.K. Infectious bovine rhinotracheitis/infectious pustular vulvovaginitis (IBR/IPV)

a I-ELISA for the detection of antibodies against BoHV-1 whole virus. Pools of up to 50 milk samples (individual or bulk milk) may be used in tests for granting and up to 100 milk samples (individual or bulk milk) may be used in tests for the maintenance of the status free from IBR/IPV.
b B-ELISA for the detection of antibodies against BoHV-1-gB protein. When referred to tests for the detection of antibodies against whole BoHV-1 in Part IV of Annex IV this method may also be used.
c B-ELISA for the detection of antibodies against BoHV-1-gE protein. Individual milk samples may be used when testing to proof the maintenance of the status free from IBR/IPV. The samples may be pooled whereat the number of samples per pool may be chosen based on documented evidence that the test is under all circumstances of day to day laboratory work sensitive enough to detect one single positive sample in the pool.
d When testing is carried out to proof the maintenance of the status free from IBR/IPV individually collected samples may be pooled. The number of samples per pool may be modulated based on documented evidence that the test system is under all circumstances of day to day laboratory work sensitive enough to detect one weak positive sample in the pool of the modulated size.
	Methods:	Matrix:
non-vaccinated bovine animals	BoHV-1 I-ELISAa	individual serum samplesd
		milk samples
	gB B-ELISAb	individual serum samplesd
		individual meat juice samples
DIVA vaccinated bovine animals with a gE-deleted vaccine	gE B-ELISAc	individual serum samples
		individual meat juice samples

Section 5 U.K. Infection with Aujeszky’s disease virus (ADV)

a ELISA for the detection of antibodies against whole ADV, ADV-gB protein or ADV-gD protein. For batch control of ADV-gB kits and ADV-gD kits or whole ADV kits, Community reference serum ADV 1, or sub-standards, must be scored positive at the dilution of 1:2. When referred to tests for the detection of whole ADV in Part V of Annex IV either of these tests may be used.
b ELISA for the detection of antibodies against ADV-gE protein. For batch control, Community reference serum ADV 1, or sub-standards, must be scored positive at the dilution 1:8.
	Methods:	Matrix:
non-vaccinated porcine animals	ADV ELISAa	individual or up to 5 pooled serum (or plasma) samples
		individual or up to 5 pooled filter paper samples
		individual meat juice samples
DIVA vaccinated porcine animals with a gE-deleted vaccine	gE ELISAb	individual serum samples

Section 6 U.K. Bovine viral diarrhoea (BVD)

1. Direct methods: U.K.

(a)

Real-time reverse transcription PCR

(b)

BVDV antigen detection ELISA

2. Serological tests: U.K.

(a)

I-ELISA

(b)

B-ELISA