Objective
To develop rapid, culture-independent tests for the prediction of drug resistances by PCR-amplification and characterizations of specific mutations within drug resistance-associated target genes
To assess risk factors contributing to MDR-TB in a case-control study
Until recently, the problem of multidrug-resistant tuberculosis (MDR-TB) in Europe was considered to be negligible, but recent data are indicative of a significant rise of MDR-TB. Several European countries might have reached a turning point of a previously benign situation. Recently, the first major MDR-TB outbreak in Europe was observed in Spain. The situation is aggravated by the uncertainty of risk factors leading to MDR-TB as well as by the long time needed for the identification of MDR by conventional techniques. To take remedial action, this project will develop rapid, culture-independent tests for the prediction of drug resistances by PCR-amplification and characterizations of specific mutations within drug resistance-associated target genes and assess risk factors contributing to MDR-TB in a case-control study.
Nested PCR protocols will be applied to amplify directly from sputum samples the drug-resistance associated target genes rpoB, inhA, katG, rrs, rpsL, 23S and gyrA which are involved in resistances to rifampicin, isoniazid, streptomycin, macrolids and fluoroquinolones. Amplification products will be directly sequenced as is already done with the PCR products from cultures. Faster mutation detection techniques - single strand conformation polymorphism (SSCP) and restriction fragment length polymorphism (RFLP), if located on a restriction site - will be tested to replace sequencing of the product as this is usually beyond the scope of a clinical diagnostic laboratory.
Fields of science
Topic(s)
Call for proposal
Data not availableFunding Scheme
CSC - Cost-sharing contractsCoordinator
80802 MUENCHEN
Germany