1. INTRODUCTION .2
Purpose3
Background3
2. TESTING STRATEGY..5
Animal selection6
Sample requirements.6
Initial screening schedule 7
Primary molecular testing with a discriminatory Western Immunoblot .8
Ring trial by additional molecular testing .9
Tertiary testing strategy 9
3. REPORTING AND REFERRAL SYSTEMS..10
Data collection and provision.11
How to refer an isolate..11
How results will be reported12
4. DISCRIMINATORY WESTERN BLOTTING METHODS13
The VLA Hybrid Western blot Method ..14
The AFSSA Discriminatory Western blot Method30
The CEA Discriminatory Test for Strain Typing of Transmissible Spongiform
Encephalopathies in Small Ruminants ..39
The FLI Method.47
The CIDC-Lelystad Discriminatory Method 57
The ISS Discriminatory Western blot.71
5. APPENDICES..87
Appendix 1: Testing Strategy Flowchart ..88
Appendix 2: Brainstem and Cerebellum Sampling Methods .89
Appendix 3: Referral Format for SR TSE Positive Isolates to the CRL STEG for
Discriminatory Testing 91
Appendix 4: List of Laboratories Participating in Primary Discriminatory and
Secondary Testing 92
Appendix 5: Summary of the Conclusions of the EFSA Opinion Regarding Appropriate
Combinations of Test and Tissue for Primary Screening .96