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Detection of antibiotic resistances of Mycobacterium tuberculosis on DNA microarrays

: Schumacher, S.; Victor, T.; Seitz, H.


International journal of infectious diseases : IJID 21 (2014), Supplement 1, S.97
ISSN: 1878-3511
ISSN: 1201-9712
International Congress on Infectious Diseases (ICID) <16, 2014, Cape Town>
Fraunhofer IBMT ()

Background: The infection with the bacterium Mycobacterium tuberculosis is one of the biggest challenges of healthcare systems, especially in South Africa or India. Associated with the extensive use of antibiotics the spread of resistances is promoted. Due to this a reliable and sensitive detection of infection and classification of resistances are needed.
Methods & Materials: DNA samples of South-African patients, provided by the Stellenbosch University, are amplified via is other-mal amplification and hybridized on a DNA microarray. The array consists of specific sequences of TB and genes involved in developing resistances against known drugs. First of all, genes leading to resistances for first line drugs lie in the focus. The immobilization of the sequences is done with a non-contact spotter that requires low amount of material. Finally a fluorescent read-out is done for a qualitative statement of infection and resistances. Advantages of this assay are e.g. a constant temperature for the isothermal amplification, avoiding heating steps and the sensitive fluorescent detection of the hybridized DNA.
Results: It is possible to detect low amounts of TB-DNA using isothermal amplification coupled with a DNA microarray. The quantity of DNA in a typical saliva or blood sample is more than sufficient. Furthermore major gene mutations responsible for resistances against first line drugs are detectable.
Conclusion: The presented method depicts a novel way to diagnose TB infection and allows a prediction about resistances to the most used antibiotics, i.e. all first line drugs. As a perspective the detection of further antibiotic resistances should be added to the assay. The explained assay has the potential to give rise to new diagnostic tools for detecting TB infection and drug resistances.