Sabine Rüsch-Gerdes National Reference Center for Mycobacteria, Forschungszentrum Borstel, Germany External proficiency panels for drug susceptibility.

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Presentation transcript:

Sabine Rüsch-Gerdes National Reference Center for Mycobacteria, Forschungszentrum Borstel, Germany External proficiency panels for drug susceptibility testing of 2 nd line drugs results from the Baltic Nordic TB-Laboratory Network

Participating laboratories Estonia, Tallin Estonia, Tartu Finland Latvia Lithuania Poland Sweden 1 Sweden 2 Germany, Borstel (Coordinator)

Drug concentrations - Borstel DrugConcentrations (µg/ml) LJ 1% BACTEC 460 MGIT 960 Amikacin Sulfate Sigma4011 Capreomycin Sulfate Sigma Cycloserine Serva40 Ethionamide Sigma2.55 Protionamide Fatol Ofloxacin Sigma222 Rifabutin Grünenthal0.5

Drug testing DrugLab 1Lab 2Lab 3Lab 4Lab 5Lab 6Lab 7Lab 8Borstel Amikacinxxxxxxxx Capreomycinxxxxxxx Clofaziminx Cycloserinxxxxxxx Ethionamide/Protionamidexxxxxxxx Kanamycinxxxx Ofloxacinxxxxxxxxx PASxxxxx Rifabutinxx Thioacetazonx

Conditions for testing I Amikacin Lab 1Lab 2Lab 3Lab 4Lab 5Lab 6Lab 7Lab 8 Substance usedAMK Disc Oxoid AMK AMK Sigma - AMK Disulfate AMK Sigma SolventADPBS-AD - DiluentADPBS-AD - Method (LJ, BACTEC etc.) BACTEC 460 Middle- brook 7H10 MGIT 960 Prop. Method - Middle- brook 7H10 BACTEC 460 Critical drug conc. (µg/ml) Capreomycin Lab 1Lab 2Lab 3Lab 4Lab 5Lab 6Lab 7Lab 8 Substance used-CM- CM Sulfate CM Sigma CM Sulfate CM Sigma Solvent-AD- AD/PBAD Diluent-AD- AD/PBAD Method (LJ, BACTEC etc.) - BACTEC 460 -MGIT 960 Prop. Method LJ Abs. Conc./ BACTEC 460 Middle- brook 7H10 BACTEC 460 Critical drug conc. (µg/ml) / AD = aqua dest. PB = phosphate buffer, PBS = phosphate buffer saline

Conditions for testing II Cycloserin Lab 1Lab 2Lab 3Lab 4Lab 5Lab 6Lab 7Lab 8 Substance used- D-CS Sigma - CS Sigma D-CS D-CS Sigma Solvent-AD0.1M PB-AD Diluent-AD- Method (LJ, BACTEC etc.) - LJ Prop. Method Middle- brook 7H10 - Prop. Method LJ Abs. Conc. LJ Critical drug conc. (µg/ml) MIC Ethionamide/ Protionamide Lab 1Lab 2Lab 3Lab 4Lab 5Lab 6Lab 7Lab 8 Substance used-ETHSigmaETHPTH FatolETH ETH Sigma Solvent- Methanol DMSO Ethylene- glycol DMSO Propylene- glycol DMSO Diluent-PBSAD AD/PBAD Method (LJ, BACTEC etc.) - BACTEC 460 Middle- brook 7H10 MGIT 960 Prop. Method LJ Abs. Conc./ BACTEC 460 LJ BACTEC 460 Critical drug conc. (µg/ml) / AD = aqua dest. PB = phosphate buffer, PBS = phosphate buffer saline

Conditions for testing III Kanamycin Lab 1Lab 2Lab 3Lab 4Lab 5Lab 6Lab 7Lab 8 Substance used---KMKM Sigma KM Sulfate -KM Sigma Solvent---AD - Diluent---AD - Method (LJ, BACTEC etc.) ---MGIT 960 Prop. Method LJ/ BACTEC BACTEC 460 Critical drug conc. (µg/ml) / Ofloxacin Lab 1Lab 2Lab 3Lab 4Lab 5Lab 6Lab 7Lab 8 Substance usedOFX Disc, Oxoid OFX OFX Sigma OFXSigma SolventADNaOH- 0.1N NaOH Propylene- glycol 0.1N NaOH Diluent-AD- PBAD Method (LJ, BACTEC etc.) BACTEC 460 BACTEC 460 Middle- brook 7H10 MGIT 960 Prop. Method BACTEC 460 LJ BACTEC 460 Critical drug concentration AD = aqua dest.

Conditions for testing IV PAS Lab 1Lab 2Lab 3Lab 4Lab 5Lab 6Lab 7Lab 8 Substance used-PASSigma-PAS -Sigma Solvent-AD - AD/PB-AD Diluent-AD - AD/PB -AD Method (LJ, BACTEC etc.) - LJ Prop. Method Middle- brook 7H10 - Prop. Method LJ/ BACTEC 460 -LJ Critical drug conc. (µg/ml) / Rifabutin Lab 1Lab 2Lab 3Lab 4Lab 5Lab 6Lab 7Lab 8 Substance used-----Farmitalia- Solvent----- Propylene- glycol - Diluent-----AD- Method (LJ, BACTEC etc.) Middle- brook 7H10 - Critical drug conc. (µg/ml) AD = aqua dest. PB = phosphate buffer

Capreomycin Strain no.Lab 1Lab 2Lab 3Lab 4Lab 5Lab 6Lab 7Lab 8Borstel 6791/02 s ssssss 3789/02 s rsssss 5922/02 s ssssss 6416/02 s ssssss 6484/02 s rsssss 6904/02 s rsssss 7040/02 s ssssss 7260/02 s ssssss 8810/02 s ssssss 9679/02 s ssssss 11019/02 s ssssss 11780/02 s rrs/rsrr 1706/03 s ssssss 1659/03 s ssssss 9647/02 s rrs/rrrr 10248/02 s ssssss 2804/02 s ssssss 8058/02 s rrs/rsrr 12555/02 s rsssrr 2032/02 s ssssss

Strain no.Lab 1Lab 2Lab 3Lab 4Lab 5Lab 6Lab 7Lab 8Borstel 6791/02 ssssssss 3789/02 rsssssss 5922/02 ssssrsss 6416/02 ssssssss 6484/02 ssssssss 6904/02 rsssssss 7040/02 ssssssss 7260/02 ssssssss 8810/02 rrrrssss 9679/02 ssssssss 11019/02 rrrsr/ssrs 11780/02 rrrsssss 1706/03 srrrrsrr 1659/03 srrssssb 9647/02 srrsssss 10248/02 srrsr/sssb 2804/02 srrrrsrr 8058/02 rrrrrsrr 12555/02 ssssssss 2032/02 ssssssss Ethionamide/Protionamide

Conclusions Further studies are necessary Equal drugs (AMK/AMK Sulfate) for testing Standardization of drug-preparation Standardized critical concentrations