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To Improve the sensitivity of the Mycobacterium tuberculosis typing technique of spoligotyping for direct application - SE3235

Spoligotyping is a DNA fingerprinting technique which is widely used in identifying and tracking strains of MTB complex organisms involved in outbreaks of disease. The technique is particularly useful for typing strains of M. bovis and is used in the surveillance programme of outbreaks of bovine tuberculosis affecting cattle and other wildlife in the UK. This PCR-based method is perfectly adequate when applied to DNA of sufficient quantity and quality, such as provided by culture but yields a partial fingerprint in up to 50% of cases when applied directly to DNA isolated from clinical samples (compared to cultures of the same cases). This may be due to the sparsity of mycobacteria in some tissues.

This proposal outlines improvements which could be made to the current method of spoligotyping to increase sensitivity of the procedure. It is hoped this will obviate the need for, or at least markedly reduce, the time of culture before typing is applied. The time to results could be reduced from a few weeks to a few days. This should find application in a range of situations, such as in routine typing of cattle tissues, slaughterhouse cases and environmental and fixed archival samples where culture may not be feasible.
Project Documents
• Final Report : SE3235 Final Report   (5111k)
Time-Scale and Cost
From: 2008

To: 2009

Cost: £58,285
Contractor / Funded Organisations
University - London - University College
Animal Diseases              
Animal Health              
Bovine Tuberculosis              
Disease Control              
Method Development              
Plants and Animals              
Fields of Study
Animal Health