P04
Update of a commercial kit for detection of Mycobacterium tuberculosis complex and a novel reagent for liquefaction and inactivation of native sputum
M Eckart(1) R Spannaus(1) V Allerheiligen(1)
1:Hain Lifescience GmbH, Hardwiesenstrasse 1, 72147 Nehren, Germany
Background
The WHO recommended FluoroType® MTBDR VER 2.0 from Hain Lifescience GmbH (a Bruker Company) is an important tool in diagnosis of TB and its resistance against first-line antibiotics rifampicin and isoniazid. A fast and sensitive detection of M. tuberculosis complex (MTBC) from native sputum is as important as an identification of resistance-mediating mutations.
Methods
The multi-copy target IS6110 was integrated into the FluoroType® MTBDR VER 2.0 as a second target for MTBC detection. An additional Liquefaction Reagent can be used to liquify and inactivate a native sputum sample in one step. Together this system enables simultaneous detection of MTBC and its associated resistance to rifampicin and isoniazid directly from native sputum specimens. The reported mutations were adapted to the recently published WHO catalogue of resistance mediating mutations.
Results
The integration of the multi-copy target IS6110 lead to an improved sensitivity of MTBC detection. The modified assay could reliably differentiate wildtype and most relevant mutations in rpoB, katG and the inhA promoter region responsible to resistance to the first-line antibiotics rifampicin and isoniazid. The Liquefaction Reagent inactivates and liquifies native sputum very effectively in one step.
Conclusions
The modified FluoroType® MTBDR VER 2.0 shows a more sensitive detection of MTBC and differentiates the most important mutations responsible for resistance based on the WHO catalogue of mutations associated with drug resistance. In combination with a novel Liquefaction Reagent it enabled sensitive detection of MTBC and identification of mutations directly from native sputum specimens.