BSI PD CEN ISO/TS 18867:2015:2016 Edition
$167.15
Microbiology of the food chain. Polymerase chain reaction (PCR) for the detection of food-borne pathogens. Detection of pathogenic Yersinia enterocolitica and Yersinia pseudotuberculosis
| Published By | Publication Date | Number of Pages |
| BSI | 2016 | 42 |
This Technical Specification specifies two horizontal methods for detection of the pathogenic bioserotypes of Y. enterocolitica and one for detection of Y. pseudotuberculosis by using real-time PCR-based methods. The described methods allow for the detection of the two pathogens in enrichments and allow the isolation of colonies. Y. pestis, the causative agent of bubonic and pneumonic plague harbours a variant of the ail gene as well and will be detected by the same primer/probe set as Y. pseudotuberculosis. However, Y. pestis is normally not associated with food. This Technical Specification is applicable to products for human consumption, animal feeding stuffs, and environmental samples.
PDF Catalog
| PDF Pages | PDF Title |
|---|---|
| 4 | Contents Page |
| 5 | European foreword |
| 8 | Foreword |
| 9 | Introduction |
| 10 | 1 Scope 2 Normative references 3 Terms and definitions 4 Principles 4.1 General |
| 11 | 4.2 Microbial enrichment 4.3 Nucleic acid extraction 4.4 Amplification and detection 4.5 Isolation 5 Reagents 5.1 General 5.2 Culture media 5.2.1 General 5.2.2 Diluent 5.2.3 Enrichment media |
| 13 | 5.2.4 Selective solid medium |
| 14 | 5.2.5 Potassium hydroxide in saline solution, KOH 5.3 Nucleic acid extraction 5.4 Reagents for PCR 5.5 Primers and probes 6 Apparatus and equipment 6.1 General |
| 15 | 6.2 Equipment for sample preparation prior to enrichment 6.3 Equipment for microbial enrichment 6.4 Equipment for nucleic acid extraction 6.5 Equipment for real-time PCR 7 Sampling 8 Procedure 8.1 Sample preparation prior to enrichment 8.1.1 General 8.1.2 Preparation of the sample |
| 16 | 8.2 Microbial enrichment 8.2.1 Pathogenic Y. enterocolitica 8.2.2 Y. pseudotuberculosis 8.2.3 Pathogenic Y. enterocolitica and Y. pseudotuberculosis 8.3 Isolation of colonies, optional 8.3.1 Pathogenic Y. enterocolitica 8.3.2 Y. pseudotuberculosis |
| 17 | 8.3.3 Process controls 8.4 Nucleic acid extraction 8.5 PCR amplification 8.5.1 General 8.5.2 PCR controls 8.6 Confirmation of the PCR product 8.6.1 General 8.6.2 Interpretation of the PCR result |
| 18 | 9 Test report |
| 19 | Annex A (normative) PCR detection and isolation of pathogenic Y. enterocolitica (see Figure A.1) |
| 20 | Annex B (informative) Real-time PCR for detection of Y. enterocolitica |
| 30 | Annex C (informative) Detection and isolation of Y. pseudotuberculosis |
| 35 | Annex D (informative) Simultaneous detection of pathogenic Y. enterocolitica and Y. pseudotuberculosis using multiplex real-time PCR |
| 38 | Bibliography |
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