EN    EN  
 EUROPEAN  
COMMISSION 
Brussels, XXX   
SANCO/11531/2013 Rev. 3 ANNEX CIS (POOL/G4/2013/11531/11531R3-
EN ANNEX CIS.doc) 
[…](2014) XXX  draft 
ANNEX 1 
  
ANNEX 
to the 
COMMISSION REGULATION (EU) No …/.. 
amending Annexe s II, VII, VIII, IX and X to Regu lation (EC) No 999/2001 of the 
European Parliament and of the Council layi ng down rules for the prevention, control 
and eradication of certain transmis sible spongiform encephalopathies 
 EN 2   EN ANNEX  
Annexes II, VII, VIII, IX and X to Regula tion (EC) No 999/2001 are amended as follows: 
(1) Annex II is amended as follows: 
(a) Points 1 and 2 of Chapter B are replaced by the following: 
"1. Structure of the risk analysis  
The risk analyses shall comprise an entry assessment and an exposure 
assessment.  
2. Entry assessment (external challenge)  
2.1. The entry assessment shall consist of assessing the likelihood that the 
BSE agent has either b een introduced into the country or region via 
commodities potentially contaminated with a BSE agent, or is already 
present in the country or region.  
The following risk factors shal l be taken into account:  
(a) the presence or absence of the BS E agent in the country or region 
and, if the agent is present, its prevalence based on the outcome of surveillance activities;  
(b) the production of meat-and-bone m eal or greaves from the BSE 
indigenous ruminant  population;  
(c) imported meat-and-bone meal or greaves;  
(d) imported bovine and ovine and caprine animals;  
(e) imported animal feed and feed ingredients;  
(f) imported products of ruminant or igin for human consumption, 
which may have contained tissues li sted in point 1 of Annex V and 
may have been fed to bovine animals;  
(g) imported products of ruminant origin for in vivo use in bovine 
animals.  
2.2. Special eradication schemes, surv eillance and other epidemiological 
investigations (especially surveilla nce for BSE conducted on the bovine 
animals population) relevant to the ri sk factors listed in point 2.1 should 
be taken into ac-count in carry ing out the entry assessment.' 
(b) In point 3 of Chapter D, Table 2 is replaced by the following: 
"Table 2 
Points targets for different adult bovine animals population sizes in a country or 
region 
Points targets for country or region 
Adult bovine animals population size 
(24 months and older) Type A 
 surveillance Type B 
 surveillance 
>1,000,000 300,000 150,000 
900,001-1,000,000 214,600 107,300 
800,001-900,000 190,700 95,350 
700,001-800,000 166,900 83,450 
600,001-700,000 143,000 71,500 EN 3   EN 500,001-600,000 119,200 59,600 
400,001-500,000 95,400 47,700 
300,001-400,000 71,500 35,750 
200,001-300,000 47,700 23,850 
100,001-200,000 22,100 11,500 
90,001-100,000 19,900 9,950 
80,001-90,000 17,700 8,850 
70,001-80,000 15,500 7,750 
60,001-70,000 13,000 6,650 
50,001-60,000 11,000 5,500 
40,001-50,000 8,800 4,400 
30,001-40,000 6,600 3,300 
20,001-30,000 4,400 2,200 
10,001-20,000 2,100 1,050 
9,001-10,000 1,900 950 
8,001-9,000 1,600 800 
7,001-8,000 1,400 700 
6,001-7,000 1,200 600 
5,001-6,000 1,000 500 
4,001-5,000 800 400 
3,001-4,000 600 300 
2,001-3,000 400 200 
1,001-2,000 200 100 
" 
(2) In Annex VII, the first paragraph in poi nt 2.2.1 of Chapter B is replaced by the 
following: 
"If BSE cannot be excluded after the resu lts of the secondary  molecular testing 
carried out in accordance with the methods  and protocols set out in Annex X, 
Chapter C, point 3.2(c) (ii), the killing and complete destruction, without delay, of all 
animals, embryos and ova identified by the inquiry referred to in  the second to fifth 
indents of point 1(b)." 
(3) In Annex VIII, Section A of Chapter A is amended as follows: 
(a) In point 1.2, paragraph (g) is replaced by the following: 
"(g) only the following ovine and caprine embryos/ova may be introduced:  
(i) embryos/ova from donor animals wh ich have been kept since birth 
in a Member State with a negligible risk of classical scrapie, or in a 
holding with a negligible or a controll ed risk of classical scrapie, or 
which meet the following requirements:  
– they are permanently identified to enable trace back to their 
holding of birth;  
– they have been kept since birth in holdings in which no case 
of classical scrapie has been confirmed during their residency; 
– they showed no clinical sign of classical scrapie at the time of 
embryo/ova collection; EN 4   EN (ii) ovine embryos/ova carryi ng at least one ARR allele." 
(b) In point 1.3, paragraph (g) is replaced by the following: 
"(g) only the following ovine and caprine embryos/ova may be introduced:  
(i) embryos/ova from donor animals wh ich have been kept since birth 
in a Member State with a negligible risk of classical scrapie, or in a 
holding with a negligible or a controll ed risk of classical scrapie, or 
which meet the following requirements:  
– they are permanently identified to enable trace back to their 
holding of birth;  
– they have been kept since birth in holdings in which no case 
of classical scrapie has been confirmed during their residency; 
– they showed no clinical sign of classical scrapie at the time of 
embryo/ova collection; 
(ii) ovine embryos/ova carryi ng at least one ARR allele."
 
(c) In point 2, the following point 3 is added: 
"2.3. The Member States or zone of the Member State with a negligible risk 
for classical scrapie are the following: 
– Austria." 
(d) Point 3.2 is replaced by the following: 
"3.2. The national scrapie control progra mmes of the following Member States 
are hereby approved:  
– Denmark, 
– Finland, 
– Sweden." 
(e) In point 4.2, paragraph (e) is  replaced by the following: 
"(e) in the case of ovine embryos, be carrying at least one ARR allele."  
(4) In Annex IX, point (ii) of point 2 of  Chapter H is replaced by the following: 
"(ii) in the case of ovine embryos, the embryos carry at least one ARR allele." 
(5) Annex X is replaced by the following: 
"ANNEX X 
REFERENCE LABORATORIES, SAMPLING AND LABORATORY 
ANALYSIS METHODS 
CHAPTER A 
National reference laboratories 
1. The designated national refe rence laboratory is to:  
(a) have at its disposal facilities and ex pert personnel enabling it to show at 
all times, and especially when the disease in question first appears, the type and strain of the agent of TSE,  and to confirm results obtained by EN 5   EN official diagnostic laboratories. Wher e it is not capable of identifying the 
strain-type of the agent, it shall se t up a procedure to ensure that the 
identification of the strain is referred to the EU reference laboratory; 
(b) verify diagnostic methods used in official diagnos tic laboratories; 
(c) be responsible for coordination of  diagnostic standards and methods 
within the Member State. To this end, it: 
– may provide diagnostic reagents to official diagnos tic laboratories; 
– is to control the quality of all diagnostic reagents used in the 
Member State; 
– is to periodically arrange comparative tests; 
– is to hold isolates of the agents  of the disease in question, or 
corresponding tissues containing su ch agents, coming from cases 
confirmed in the Member State; 
– is to ensure confirmation of results obtained in diagnostic 
laboratories; 
(d) is to cooperate with the EU reference laboratory, which includes the 
participation in the periodic comp arative tests organised by the EU 
reference laboratory. Should a nationa l reference laboratory fail in a 
comparative test organised by the EU reference laboratory, it shall take 
immediately all the corrective actions to remedy the situation and successfully pass the repeat comparativ e test or the next comparative test 
organised by the EU reference laboratory.  
2. However, by way of derogation from poi nt 1, Member States which do not 
have a national reference laboratory shall use the services of the EU reference 
laboratory or of national reference laborat ories located in other Member States 
or European Free Trade Asso ciation (EFTA) Members. 
3. The national referen ce laboratories are:  
Austria:  Austrian Agency for Health and Food Safety- AGES 
Institute for Veterinary Disease Control Robert Koch Gasse 17 A-2340 Mödling 
Belgium: CERVA-CODA-VAR 
Centre d'Étude et de Recherches Vétérinaires et Agrochimiques, Centrum voor Onderzoek in Diergeneeskunde en Agrochemie, Veterinary and Agrochemical Research Centre Groeselenberg 99 B-1180 Bruxelles 
Bulgaria: Национален  диагностичен  научноизследователски  
ветеринарномедицински  институт  ‘Проф . Д-р Георги  
Павлов ’ 
Национална  референтна  лаборатория  ‘Tрансмисивни  
спонгиформни  енцефалопатии ’ 
бул. ‘
Пенчо  Славейков ’ 15 
София  1606 EN 6   EN (National Diagnostic Veterinary  Research Institute ‘Prof. 
Dr. Georgi Pavlov’, National Reference Laboratory for 
Transmissible Spongiform Encephalopathies, 15 Pencho Slaveykov Blvd., 1606 Sofia) 
Croatia Croatian Veterinary Institute, 
Savska Cesta 143 10000 Zagreb 
Cyprus: State Veterinary Laboratories 
Veterinary Services CY-1417 Athalassa Nicosia 
Czech Republic: Státní vete rinární ústav Jihlava (State Veterinary Institute 
Jihlava) National Reference Laboratory for BSE and Animal TSEs Rantířovská 93 
586 05 Jihlava 
Denmark: Veterinærinstituttet 
Danmarks Tekniske Universitet Bülowsvej 27 DK-1870 Frederiksberg C (National Veterinary Institute, Technical University of Denmark, 27, Bülowsvej, DK - 1870 Frederiksberg C) 
Estonia: Veterinaar- ja Toidulab oratoorium (Estonian Veterinary 
and Food Laboratory ) Kreutzwaldi 30 Tartu 51006 
Finland: Finnish Food Safety Authority Evira 
Research and Laboratory Department Veterinary Virology Re search Unit- TSEs 
Mustialankatu 3 FI-00790 Helsinki 
France: ANSES-Lyon, Unité MND 
31, avenue Tony Garnier 69 364 LYON CEDEX 07 
Germany: Friedrich-Loeffler-Institut 
Institute for Novel and Emerging Infectious Diseases at the Friederich-Loeffler-Institut Federal Research Institute for Animal Health Suedufer 10 D-17493 Greifswald Insel Riems 
Greece: Ministry of Agriculture — Veterinary Laboratory of 
Larissa 6th km of Larissa — Trikala Highway 
GR-41110 Larissa 
Hungary: Veterinary Diagnostic Di rectorate, National Food Chain 
Safety Office (VDD NFCSO) EN 7   EN Tábornok u. 2 
1143 Budapest 
Ireland: Central Veterinary Research Laboratory 
Department of Agriculture, Food and the Marine Backweston Campus Celbridge Co. Kildare 
Italy: Istituto Zooprofilattic o Sperimentale del Piemonte, 
Liguria e Valle d’Aosta — CEA Via Bologna, 148 I-10154 Torino 
Latvia: Institute of Food Safety, Animal Health and Environment 
(BIOR) Lejupes Str. 3 Riga LV 1076 
Lithuania: National Food and Veteri nary Risk Assessment Institute 
J. Kairiūkščio str. 10 
LT-08409 Vilnius 
Luxembourg: CERVA-CODA-VAR 
Centre d'Étude et de Recherches Vétérinaires et Agrochimiques, Centrum voor Onderzoek in Diergeneeskunde en Agrochemie, Veterinary and Agrochemical Research Centre Groeselenberg 99 B-1180 Bruxelles 
Malta: Veterinary Diagnostic Laboratory 
Department of Food Health and Diagnostics 
Veterinary Affairs and Fisheries Division Ministry for Rural Affairs and the Environment  Albert Town Marsa 
Netherlands: Central Veterinary Instutute of Wageningen UR  
Edelhertweg 15 8219 PH Lelystad P.O. Box 2004 NL-8203 AA Lelystad 
Poland: Pa ństwowy Instytut Weterynaryjny (PIWet) 
24-100 Pu ławy 
al. Partyzantów 57 
Portugal: Setor diagnóstico EET 
Laboratório de Patologia Unidade Estratégica de Investigação e Serviços de Produção e Saúde Animal Instituto Nacional de Investig ação Agrária e Veterinária 
Rua General Morais Sarmento 1500-311 Lisboa 
Romania: Institutul de Diagnostic și Sănătate Animal ă (Institute for EN 8   EN Diagnosis and Animal Health) 
Department of Morphology Strada Dr. Staicovici nr. 63, 5 
București 050557 
Slovakia: State Veterinary Institute Zvolen 
Pod dráhami 918 SK-960 86, Zvolen 
Slovenia: University of Lj ubljana, Veterinary faculty 
National Veterinary Institute Gerbičeva 60 
SI-1000 Ljubljana 
Spain: Laboratorio Central de Veterinaria (Algete) 
Ctra. M-106 pk 1,4 28110 Algete (Madrid) 
Sweden: National Veterinary Institute 
S-751 89 Uppsala 
United Kingdom: Animal Health a nd Veterinary Laboratories Agency 
Woodham Lane New Haw, Addlestone, Surrey KT15 3NB 
CHAPTER B 
EU reference laboratory 
1. The EU reference laboratory for TSEs is:  
The Animal Health and Veterinary Laboratories Agency  
Woodham Lane  New Haw  Addlestone  
Surrey KT15 3NB  
United Kingdom 
2. The functions and duties of the EU reference laboratory are: 
(a) to coordinate, in consultation with the Commission, the methods 
employed in the Member States for diagnosing TSEs and the determination of the prion protein genot ype in ovine animals, specifically 
by: 
– storing and supplying correspondi ng tissues containing the TSE 
agents, for the development or production of the relevant diagnostic tests or for typing strains of the TSE agents; 
– supplying standard sera and other re ference reagents to the national 
reference laboratories in order to standardise the tests and reagents 
used in the Member States; 
– building up and retaining a co llection of corresponding tissues 
containing the agents and strains of TSEs; EN 9   EN – organising periodic comparative te sts for the procedures for the 
diagnosis of TSEs and for the determination of the prion protein genotype in ovine animals at EU level; 
– collecting and collating data and information on the methods of 
diagnosis used and the results of  tests carried out in the EU; 
– characterising isolates of the TSE agent by the most up-to-date 
methods to allow greater understand ing of the epidemiology of the 
disease; 
– keeping abreast of trends in surveillance, epidemiology and 
prevention of TSEs throughout the world; 
– maintaining expertise on prion dise ases to enable rapid differential 
diagnosis; 
– acquiring a thorough knowledge of the preparation and use of 
diagnostic methods used to control and eradicate TSEs; 
(b) to assist actively in the diagnosis of outbreaks of TSEs in Member States 
by studying samples from TSE-infected  animals sent for confirmatory 
diagnosis, characterisation a nd epidemiological studies; 
(c) to facilitate the training or retraini ng of experts in laboratory diagnosis 
with a view to the harmonisation of  diagnostic techniques throughout the 
EU. 
CHAPTER C 
Sampling and laboratory testing 
1. Sampling  
Any samples intended to be examined for the presence of a TSE shall be 
collected using the methods and protocols laid down in the latest edition of the Manual for diagnostic tests and vaccines for Terrestrial Animals of the World 
Organisation for Animal Health (OIE) (t he Manual). In addition to, or in the 
absence of, OIE methods and protocols, and to ensure that sufficient material is 
available, the competent authority sha ll ensure the use of sampling methods 
and protocols in accordance with guide lines issued by the EU reference 
laboratory. 
In particular the competent authority shall collect the appropriate tissues, 
according to the available scientific advice and the guidelines of the EU reference laboratory, in or der to ensure the detectio n of all known strains of 
TSE in small ruminants and shall keep at least half of the collected tissues fresh but not frozen until the result of the ra pid test is negative. Where the result is 
positive or inconclusive the residual tissues must be subject to confirmatory testing, and be processed subsequently in accordance with the EU reference 
laboratory guidelines on discriminatory te sting and classification - "TSE strain 
characterisation in small ruminant s: A technical handbook for National 
Reference Laboratories in the EU". 
The samples shall be correctly marked as to the identity of the sampled animal. 
2. Laboratories  EN 10   EN Any laboratory examination for TSE shall be carried out in official diagnostic 
laboratories designated for that pur pose by the competent authority. 
3. Methods and protocols 3.1. Laboratory testing for the pres ence of BSE in bovine animals 
(a) Suspect cases  
Samples from bovine animals sent for laboratory testing pursuant to the 
provisions of Article 12(2) sha ll immediately be subjected to 
confirmatory examinations using at least one of the following methods and protocols laid down in the latest edition of the Manual: 
(i) the immunohistochemical (IHC) method; (ii) Western blot; (iii) the demonstration of characteris tic fibrils by electron microscopy;  
(iv) histopathological examination;  (v) the combination of rapid tests as laid down in the third 
subparagraph. 
If the histopathological examination is inconclusive or negative, the 
tissues shall be submitted to a furt her examination by one of the other 
confirmatory methods and protocols.  
Rapid tests may be used for both prim ary screening of suspect cases and, 
if inconclusive or positive, for s ubsequent confirmation, according to the 
guidelines from the EU reference labora tory - "OIE rules for the official 
confirmation of BSE in bovines (based on an initial reac tive result in an 
approved rapid test) by using a seco nd rapid test", and provided that: 
(i) the confirmation is carried out in  a national reference laboratory for 
TSEs; and  
(ii) one of the two rapid tests is a Western blot; and  
(iii) the second rapid test used:  
– includes a negative tissue cont rol and a bovine BSE sample 
as positive tissue control,  
– is of a different type than the test used for the primary 
screening; and  
(iv) if a rapid Western blot is used as the first test, the result of that test 
must be documented and the blot image submitted to the national reference laboratory for TSEs; and  
(v) where the result of the primary screening is not confirmed by the 
subsequent rapid test, the samp le must be subjected to an 
examination by one of the other confirmatory methods; where the 
histopathological examination is used for that purpose, but proves to be inconclusive or negative, the tissues must be submitted to a further examination by one of the other confirmatory methods and 
protocols. EN 11   EN If the result of one of the confirmato ry examinations referred to in points 
(i) to (v) of the first subparagraph is positive, the animal shall be regarded as a positive BSE case. 
(b) BSE monitoring  
Samples from bovine animals sent for laboratory testing pursuant to the 
provisions of Annex III, Chapter A, Pa rt I shall be examined by a rapid 
test. 
When the result of the rapid test is inconclusive or positive, the sample 
shall immediately be subjected to c onfirmatory examinations using at 
least one of the following methods a nd protocols laid down in the latest 
edition of the Manual: 
(i) the immunohistochemical (IHC) method;  (ii) Western blot;  (iii) the demonstration of characteris tic fibrils by electron microscopy;  
(iv) histopathological examination;  (v) the combination of rapid tests as laid down in the fourth 
subparagraph.  
Where the histopathological examinati on is inconclusive or negative, the 
tissues shall be submitted to a furt her examination by one of the other 
confirmatory methods and protocols.  
Rapid tests may be used for both prim ary screening and, if inconclusive 
or positive, for subsequent confirmation, according to the guidelines from the EU reference laboratory - "OIE rules for the official confirmation of BSE in bovines (based on an initial reactive result in an approved rapid 
test) by using a second rapid test", and provided that: 
(i) the confirmation is carried out in  a national reference laboratory for 
TSEs; and 
(ii) one of the two rapid tests is a Western blot; and  (iii) the second rapid test used:  
– includes a negative tissue cont rol and a bovine BSE sample 
as positive tissue control,  
– is of a different type than the test used for the primary 
screening; and  
(iv) if a rapid Western blot is used as the first test, the result of that test 
must be documented and the blot image submitted to the national reference laboratory for TSEs; and  
(v) where the result of the primary screening is not confirmed by the 
subsequent rapid test, the samp le must be subjected to an 
examination by one of the other confirmatory methods; where the 
histopathological examination is used for that purpose, but proves to be inconclusive or negative, the tissues must be submitted to a further examination by one of the other confirmatory methods and 
protocols.  EN 12   EN An animal shall be regarded a positive BSE case if the result of the rapid 
test is inconclusive or positive, and at least one of the confirmatory examinations referred to in points (i) to (v) of the second subparagraph is 
positive. 
(c) Further examination of positive BSE cases 
Samples from all positive BSE cases sh all be forwarded to a laboratory, 
appointed by the competent aut hority, which has participated 
successfully in the latest proficiency testing organised by the EU reference laboratory for discriminatory testing of confirmed BSE cases, 
where they shall be further tested in accordance with the methods and 
protocols laid down in the EU refe rence laboratory's method for the 
classification of bovine TSE isolat es (a two-blot method for the 
provisional classification of bovine TSE isolates). 
3.2. Laboratory testing for the presence of  TSE in ovine and caprine animals 
(a) Suspect cases 
Samples from ovine and caprine anim als sent for laboratory testing 
pursuant to the provisions of Article 12(2) shall immediately be subjected 
to confirmatory examinations using at least one of the following methods 
and protocols laid down in the la test edition of the Manual:  
(i) the immunohistochemical (IHC) method;  (ii) Western blot;  (iii) the demonstration of characteris tic fibrils by electron microscopy;  
(iv) histopathological examination.  In case the histopathological examinati on is inconclusive or negative, the 
tissues shall be submitted to a furt her examination by one of the other 
confirmatory methods and protocols.  
Rapid tests may be used for primary screening of suspect cases. Such 
tests may not be used for subsequent confirmation. 
Where the result of the rapid test us ed for primary screening of suspect 
cases is positive or inconclusive, the sample shall be subjected to an examination by one of the confirmatory examinations referred to in points (i) to (iv) of the first subpa ragraph. Where the histopathological 
examination is used for that purpose, but proves to be inconclusive or negative, the tissues shall be submitted to a further examination by one of the other confirmatory methods and protocols.  
If the result of one of the confirmato ry examinations referred to in points 
(i) to (iv) of the firs t subparagraph is positive, the animal shall be 
regarded as a positive TSE case and further examination as referred to in point (c) shall be performed.  
(b) TSE monitoring   
Samples from ovine and caprine anim als sent for laboratory testing 
pursuant to the provisions of Annex I II, Chapter A, Part II (Monitoring in 
ovine and caprine animals)  shall be examined by a rapid test, in order to 
ensure the detection of a ll known strains of TSE.  EN 13   EN When the result of the rapid test is inconclusive or positive, the sampled 
tissues shall immediately be sent to an official laboratory for 
confirmatory examinations by hi stopathology, immunohistochemistry, 
Western blotting or demonstration of  characteristic fibrils by electron 
microscopy, as referred to in point (a).  If the result of the confirmatory 
examination is negative or inconclusive, the tissues shall be submitted to a further examination by immunohist ochemistry or Western blotting. 
If the result of one of the confirmatory examinations is positive, the 
animal shall be regarded as a positive TSE case and further examination as referred to in point  (c) shall be performed. 
(c) Further examination of positive TSE cases 
(i) Primary molecular testing with a discriminatory Western blotting 
method 
Samples from clinical suspect cases and from animals tested in 
accordance with Annex III, Chapte r A, Part II, points 2 and 3 
which are regarded as positive TSE cases but which are not atypical scrapie cases following the examinations referred to in 
points (a) or (b), or which displa y characteristics which are deemed 
by the testing laboratory to merit investigation, shall be examined 
using a discriminatory Wester n blotting method listed in the 
guidelines of the EU reference labo ratory by an official diagnostic 
laboratory designated by the co mpetent authority, which has 
participated successfully in the latest proficiency testing organised by the EU reference laboratory for the use of such a method.  
(ii) Secondary molecular testing with additional molecular testing 
methods 
TSE cases in which the presence of BSE cannot be excluded 
according to the guidelines issued by the EU reference laboratory by the primary molecular testing refe rred to in point (i), shall be 
referred immediately to the EU reference laboratory, with all the relevant information available. The samples shall be submitted to further investigation and confirmation by at least one alternative method, differing immunochemically from the original primary molecular method, as described in the guidelines of the EU reference laboratory depending on the volume and nature of the 
referred material. These additional tests will be carried out in the following laboratories approved for the relevant method: 
Agence Nationale de Sécurité Sanitaire de l’alimentation, de 
l’environnement et du travail  31, avenue Tony Garnier  BP 7033  F-69342 Lyon Cedex 
Commissariat à l’Energie Atomique 
18, route du Panorama BP 6  F-92265 Fontenay-aux-Roses Cedex  
Animal Health and Veterinary Laboratories Agency  EN 14   EN Woodham Lane  
New Haw  Addlestone  Surrey KT15 3NB  United Kingdom  
The results shall be interpreted by the EU reference laboratory 
assisted by a panel of experts re ferred to as the Strain Typing 
Expert Group (STEG), including a representative of the relevant 
national reference laboratory. The Commission shall be informed 
immediately about the outcom e of that interpretation. 
(iii) Mouse bioassay 
Samples indicative of BSE or inconclusive for BSE, following 
secondary molecular testing, shal l be further analysed by mouse 
bioassay for final confirmation. The na ture or quantity of available 
material may influence the bioassay design, which will be approved by the EU reference laboratory assisted by the STEG on a case by case basis. Bioassays will be performed by the EU reference laboratory, or by laboratories designated by the EU reference 
laboratory. 
The results shall be interpreted by the EU reference laboratory 
assisted by the STEG. The Co mmission shall be informed 
immediately about the outcom e of that interpretation. 
3.3. Laboratory testing for the presence of TS Es in species other than those referred 
to in points 3.1 and 3.2 
Where methods and protocols are establis hed for tests carried out to confirm 
the suspected presence of a TSE in a species other than bovine, ovine and 
caprine, they shall include at least a histopathological examination of brain 
tissue. The competent authority may also  require laboratory tests such as 
immunohistochemistry, Western blotting, demonstration of characteristic 
fibrils by electron microscopy or other me thods designed to detect the disease 
associated form of the prion protein. In  any case at least one other laboratory 
examination shall be carried out if the initial histopathological examination is negative or inconclusive. At least three different examinations with positive results shall be carried out in the event of  the first appearance of the disease.  
In particular, where BSE is suspected in  a species other than bovine animals, 
the cases shall be referred to the EU re ference laboratory assisted by a panel of 
experts referred to as the Strain Typing Expert Group (STEG) for further 
characterisation.  
4. Rapid tests  
For the purposes of carrying out the rapid tests in accordance with Articles 5(3) 
and 6(1), only the following methods sh all be used as rapid tests for the 
monitoring of BSE in bovine animals: 
– the immunoblotting test based on a Western blotting procedure for the 
detection of the Proteinase K-resist ant fragment PrPRes (Prionics-Check 
Western test), EN 15   EN – the sandwich immunoassay for PrPRes  detection (short assay protocol) 
carried out following denaturation and concentration steps (Bio-Rad TeSeE SAP rapid test), 
– the microplate-based immunoassay (ELI SA) which detects Proteinase K-
resistant PrPRes with monoclonal an tibodies (Prionics-Check LIA test), 
– the immunoassay using a chemical pol ymer for selective PrPSc capture 
and a monoclonal detection antibody di rected against conserved regions 
of the PrP molecule (IDEXX HerdChek BSE Antigen Test Kit, EIA & HerdChek BSE-Scrapie Antigen (IDEXX Laboratories)), 
– the lateral-flow immunoassay using two different monoclonal antibodies 
to detect Proteinase K-resistan t PrP fractions (Prionics Check 
PrioSTRIP), 
– the two-sided immunoassay using tw o different monoclonal antibodies 
directed against two epitopes presen ted in a highly unfolded state of 
bovine PrPSc (Roboscreen Beta Prion BSE EIA Test Kit). 
For the purposes of carrying out the rapid tests in accordance with Articles 5(3) 
and 6(1), only the following methods sh all be used as rapid tests for the 
monitoring of TSE in ovin e and caprine animals:  
– the sandwich immunoassay for PrPRes  detection (short assay protocol) 
carried out following denaturation and concentration steps (Bio-Rad TeSeE SAP rapid test), 
– the sandwich immunoassay for PrPR es detection with the TeSeE 
Sheep/Goat Detection kit carried out following denaturation and concentration steps with the TeSeE Sheep/Goat Purification kit (Bio-Rad 
TeSeE Sheep/Goat rapid test), 
– the immunoassay using a chemical pol ymer for selective PrPSc capture 
and a monoclonal detection antibody di rected against conserved regions 
of the PrP molecule (HerdChe k BSE-Scrapie Antigen (IDEXX 
Laboratories)), 
– the lateral-flow immunoassay using two different monoclonal antibodies 
to detect Proteinase K-resistan t PrP fractions (Prionics - Check 
PrioSTRIP SR, visual reading protocol).  
In all rapid tests, sample tissue on which the test must be applied must comply 
with the manufacturer’s in structions for use.  
Producers of rapid tests must have a quality assurance system in place that has 
been approved by the EU reference la boratory and ensures that the test 
performance does not change. Producer s must provide the EU reference 
laboratory with the test protocols.  
Changes to rapid tests and to test pr otocols may only be made after prior 
notification to the EU reference laborato ry and provided that the EU reference 
laboratory finds that the change does not  alter the sensitivity, specificity or 
reliability of the rapid test. That finding shall be communicated to the Commission and to the national reference laboratories.  
5. Alternative tests  EN 16   EN (To be defined)"