EURL
European Union Reference Laboratory for
Brucellosis
Methods
This document is applicable to Bison ssp, Bos ssp and Bubalus ssp as listed in the Commission Implementing Regulation (EU) 2018/1882 and describes the guidelines relative to method validation and testing strategy in these species.
Specific guidelines for Bubalus ssp. are in preparation.
According to the Commission Delegated Regulation (EU) 2020/689 Article 6, specific legislation and guidance for brucellosis testing will be made available:
(a) in the websites of the EURL and COM: see Methods | EURL (anses.fr) ;
(b) if no recommendation is provided by EURL websites, OIE Manuals have to be applied
(c) if no recommendation is provided by EURL / OIE websites, Article 34 of Regulation (EU) 2017/625 should be applied:
• National rules. In absence of national rules, relevant methods recommended by NRL validated in accordance with international standards or
• Relevant methods validated with inter or intra laboratory methods validation studies
• In case of urgency, NRL (in absence, other designated lab) may use methods which have not been validated
The present document describes the requirements for validation (fitness for purpose) of antigen for Rose Bengal test (RBT) in brucellosis diagnostic.
The standardisation and validation are critical steps when a method is intended for routine diagnostic use in multiple laboratories. Therefore, customers can have confidence in the results produced by the test. All diagnostic assays should be validated for the species which they will be used for, according to the new OIE and EU principles and methods.
To validate a method, the manufacturer must submit a file to the National Reference Laboratory for Brucellosis (NRL) including a descriptive administrative part of the reagent(s) and a technical part. The evaluation criteria are described in the downloadable document (3. Criteria for Rose Bengal Antigen validation). The dossier will be reviewed by the NRL and a report will be returned to the manufacturer (Annex A).
The present document describes the requirements for validation (fitness for purpose) of antigen for complement fixation test (CFT) in brucellosis diagnostic. The standardisation and validation are critical steps when a method is intended for routine diagnostic use in multiple laboratories. Therefore, customers can have confidence in the results produced by the test. All diagnostic assays should be validated for the species which they will be used for, according to the new OIE and EU principles and methods.
To validate a method, the manufacturer must submit a file to the National Reference Laboratory for Brucellosis (NRL) including a descriptive administrative part of the reagent(s) and a technical part. The evaluation criteria are described in the downloadable method (3. Criteria for Complement Fixation Antigen validation). The dossier will be reviewed by the NRL and a report will be returned to the manufacturer.
The Brucellin Skin Test (BST) is a cell-mediated immune assay which is based on the use of purified and standardised protein allergen, almost completely devoid of lipopolysaccharide. The BST relies on the delayed-type hypersensitivity reaction whose results are interpreted 48 and 72 hours post brucellin injection in small Ruminants and Cattle respectively.
Therefore an animal is restrained twice: to administer brucellin, and to read the result.
The present document describes a standard technique aiming at controlling the fulfilment of OIE and EU requirements regarding the standardisation of indirect enzyme-linked immunosorbent assays (IELISA) kits for the detection of antibodies specific to smooth Brucella species (especially B. abortus, B. melitensis and B. suis) in bovine tank milk.
(cattle only)
The present document describes a technique aiming at detecting antibodies specific of smooth Brucella species (B. abortus, B. melitensis and B. suis) in bovine milk from the bulk tank by the Milk Ring-Test (MRT).
In lactating cattle, the MRT can be used for screening herds for brucellosis. In large herds (> 100 lactating cows), the sensitivity of the test becomes less reliable.
False-positive reactions may occur in cattle vaccinated less than 4 months prior to testing, in samples containing abnormal milk (such as colostrum) or in cases of mastitis.
The present document describes the requirements for validation (fitness for purpose) of ELISA kits for brucellosis diagnostic. The fulfilment of the OIE and EU requirements regarding the validation of indirect and competitive enzyme-linked immunosorbent assays (I-ELISA and C-ELISA) kits for the detection of antibodies specific to smooth Brucella species (especially B. abortus, B. melitensis and B. suis) in sera/milk samples from various animal species. The standardisation and validation are critical steps when a method is intended for routine diagnostic use in multiple laboratories. Therefore, customers can have confidence in the results produced by the test. All diagnostic assays should be validated for the species which they will be used for, according to the new OIE and EU principles and methods. For validation, the manufacturer must submit a file to the National Reference Laboratory for Brucellosis (NRL) including a descriptive administrative part of the reagent(s) and a technical part. The evaluation criteria are described in the document (3. Criteria for ELISA kits validation).
The present document describes a standard technique aiming at controlling the fulfilment of OIE and EU requirements regarding the standardisation of indirect enzyme-linked immunosorbent assays (iELISA) kits for the detection of antibodies specific to smooth Brucella species (especially B. abortus, B. melitensis and B. suis) in bovine individual sera or pools of sera.
The present document describes a standard technique aiming at controlling the fulfilment of OIE and EU requirements regarding the standardisation of diagnostic antigens for the detection of antibodies specific of smooth Brucella species (especially B. abortus, B. melitensis and B. suis):
- in animal individual sera by the Rose Bengal Test (RBT) and the Complement Fixation Test (CFT);
- in bovine pooled milk samples by the Milk Ring-Test (MRT).
The present document describes a standard technique aiming at detecting antibodies specific of Brucella ovis by the complement fixation test in ovine sera.