Method for the detection of bacterial species of the genera anaplasma/ehrlichia and bartonella

Abstract

The present invention relates to a method for the detection and identification of bacterial species belonging to the genera Anaplasma / Ehrlichia and Bartonella, and also provides triggers and probes required for its application, as well as associated kits.

Description

[0001]This application claims the benefit of priority to Spanish Patent Application No. ES 200701830, filed Jun. 29, 2007, which is hereby incorporated by reference in its entirety.FIELD OF THE INVENTION[0002]The present invention relates to a method for the detection and identification of bacterial species belonging to the genera Anaplasma / Ehrlichia and Bartonella (ES 2264642), and also provides triggers and probes required for its application, as well as associated kits.BACKGROUND[0003]To date, about 200 zoonotic diseases (e.g., bartonelosis, leptospirosis, Lyme borreliosis, etc.), which affect humans and represent one of the main causes of death and entail substantial economic loss in third world countries, have been described. Coexistence with animals, lack of sanitary infrastructure and low cultural level continue to be the main allies of these diseases.[0004]In the same manner, certain types of zoonosis which are widespread in third world countries are now thriving in industri...

AI Technical Summary

Benefits of technology

[0010]The present invention relates to the use of genes 16S and msp2 and intergenic space 16S-23S for the detection and identification of the different bacterial species and groups of species that cause zoonosis, improving the previously described procedures based on its capacity to specifically detect a substantial number of bacterial species using probes and triggers with high sensitivity levels.

Problems solved by technology

These circumstances, amongst others, entail the risk of introducing exotic diseases into the environment.

Said medical profiles, due to their high degree of similarity, do not allow a fast and reliable identification of the pathogenous agent, so that specific and fast treatment is not possible and is occasionally administered too late.

To date, the molecular diagnosis methods available are basically limited to the detection of pathogens based on antibody technology.

These types of techniques are scarcely applicable to certain species of genera, such as Bartonella and Anaplasma / Ehrlichia, due to the fact that said species do not normally grow in regular culture mediums and may even require cellular cultures.

Said technology, while being highly effective, is greatly limited by the difficulty in finding specific markers or regions, in addition to triggers and probes, which ensure reliable sample analysis.

Other methods, such as those described by U.S. Pat. Nos. 6,300,072 and 6,518,020, are capable of detecting and identifying bacteria of the genus Bartonella by using the same region (16S-23S), even though the number of species within this genus has increased substantially since said patents were filed and their approximation, which consists of discriminating between species according to the size of the amplicon obtained during PCR, is not useful for certain known species within the same genus which are similar in size to the amplified fragment.

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