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Development of a Sensitive and Specific Molecular Typing Method for the Epidemiological Study of Salmonella - OZ0312

The objective of this proposal work is to establish a system that will allow the objective genetic identification and comparison of Salmonella spp. thereby improving strain identification. The technology will include the use of highly reproducible AFLP DNA profiling and ABI fragment analysis software to form a database of profiles and markers of specific traits. The establishment of such a system would allow the easy comparison of strains isolated from the various biological (human or animal), food and environmental sources at various locations in the UK. This has the potential to significantly enhance the sharing of informatioon and the tracing of foodbourne pathogen outbreaks on a national scale. The DNA profiling data obtained during the course of this study will form the framework of the database which will need to be expanded and the system further validated by multi-laboratory trials.
The overall objective of this proposal is to carry out analysis to identify the polymorphic DNA markers that will reproducibly represent specific traits in foodbourne Salmonella strains thereby establishing and assessing the framework of a working database.

1) Identify and catalogue representative strains with respect to such known characteristics as geographical origin, time of isolation, biological/ environmental origin, clinical manifestations, toxin production (heat stable or heat lable), phage type, PFGE profile and serogroup.

2) Produce, through empirical determination, a robust, reproducible AFLP protocol with well defined limitations.

3) Maximise sensitivity of the AFLP technique (with respect to distinguishing closely related strains by identifying unique polymorphic makers.)

4) Identify reproducible polymorphic DNA markers and by comparison with known characteristics and statistical analysis assign specific traits to markers and define identification categories for database framework.

5) Challenge and assess system by analysis of a minimum of 40 blind sample. Draw comparisons with currently accepted methodologies employed in typing this organism
Project Documents
• Final Report : Development of a sensitive and specific molecular typing method for the epidemiological study of Salmonella   (932k)
Time-Scale and Cost
From: 2000

To: 2003

Cost: £228,700
Contractor / Funded Organisations
Animal Health              
GM Non-Food              
Plants and Animals              
Fields of Study
Animal Health