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Council Directive 98/57/EC of 20 July 1998 on the control of Ralstonia solanacearum (Smith) Yabuuchi et al.
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General principles | |||||
SECTION I: | Application of the test scheme | ||||
1. | Detection scheme for the diagnosis of brown rot and bacterial wilt ( R. solanacearum ) in potato tubers and potato, tomato or other host plants with symptoms of brown rot or bacterial wilt | ||||
2. | Scheme for detection and identification of R. solanacearum in samples of asymptomatic potato tubers | ||||
3. | Scheme for detection and identification of R. solanacearum in samples of asymptomatic, potato, tomato or other host plants | ||||
SECTION II: | Detailed methods for detection of R. solanacearum in potato tubers and potato, tomato or other host plants with symptoms of brown rot or bacterial wilt | ||||
1. | Symptoms | ||||
2. | Rapid screening tests | ||||
3. | Isolation procedure | ||||
4. | Identification tests for R. solanacearum | ||||
SECTION III: | 1. | Detailed methods for detection and identification of R. solanacearum in samples of asymptomatic potato tubers | |||
1.1. | Sample preparation | ||||
1.2. | Testing | ||||
2. | Detailed methods for detection and identification of R. solanacearum in samples of asymptomatic potato, tomato or other plants | ||||
2.1. | Sample preparation | ||||
2.2. | Testing | ||||
SECTION IV: | 1. | Scheme for detection and identification of R. solanacearum in water | |||
2. | Methods for detection and identification of R. solanacearum in water | ||||
2.1. | Sample preparation | ||||
2.2. | Testing | ||||
SECTION V: | 1. | Scheme for detection and identification of R. solanacearum in soil | |||
2. | Methods for detection and identification of R. solanacearum in soil | ||||
2.1. | Samples preparation | ||||
2.2. | Testing | ||||
SECTION VI: | Optimised protocols for detection and identification of R. solanacearum | ||||
A | Diagnostic and detection tests | ||||
1. | Stem streaming test | ||||
2. | Detection of poly-ß-hydroxybutyrate granules | ||||
3. | Serological agglutination tests | ||||
4. | Selective isolation | ||||
4.1. | Selective plating | ||||
4.2. | Enrichment procedure | ||||
5. | Immunofluorescence test (IF test) | ||||
6. | Polymerase chain reaction test (PCR test) | ||||
6.1. | DNA purification methods | ||||
(a) | Method according to Pastrik (2000) | ||||
(b) | Other methods | ||||
6.2. | PCR | ||||
6.3. | Analysis of the PCR product | ||||
7. | Fluorescent in-situ hybridization test (FISH test) | ||||
8. | Enzyme Linked ImmunoSorbent Assay (ELISA) tests | ||||
(a) | Indirect ELISA | ||||
(b) | DASI (Double-Antibody Sandwich Indirect) ELISA | ||||
9. | Bioassay test | ||||
B | Identification tests | ||||
1. | Nutritional and enzymatic identification tests | ||||
2. | IF test | ||||
3. | ELISA test | ||||
4. | PCR test | ||||
5. | FISH test | ||||
6. | Fatty acid profiling (FAP) | ||||
7. | Strain characterisation methods | ||||
7.1. | Biovar determination | ||||
7.2. | Genomic fingerprinting | ||||
7.3. | PCR methods | ||||
C | Confirmation test | ||||
Appendix 1 | Laboratories involved in optimisation and validation of protocols | ||||
Appendix 2 | Media for isolation and culture of R. solanacearum | ||||
Appendix 3 | (A) | Commercially available standardised control material | |||
(B) | Preparation of controls | ||||
Appendix 4 | Buffers for test procedures | ||||
Appendix 5 | Determination of contamination level in IF and FISH tests | ||||
Appendix 6 | Validated PCR protocols and reagents | ||||
Appendix 7 | Validated reagents for FISH test | ||||
Appendix 8 | Culture conditions for tomato and eggplant | ||||
References | ] |
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