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

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Presentation on theme: "Sabine Rüsch-Gerdes National Reference Center for Mycobacteria, Forschungszentrum Borstel, Germany External proficiency panels for drug susceptibility."— Presentation transcript:

1 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

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

3 Drug concentrations - Borstel DrugConcentrations (µg/ml) LJ 1% BACTEC 460 MGIT 960 Amikacin Sulfate Sigma4011 Capreomycin Sulfate Sigma 401.252.5 Cycloserine Serva40 Ethionamide Sigma2.55 Protionamide Fatol401.252.5 Ofloxacin Sigma222 Rifabutin Grünenthal0.5

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

5 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) 1.04.06.01.0 -8.01.0 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) -10.0-1.252.520.0/5.02.01.25 AD = aqua dest. PB = phosphate buffer, PBS = phosphate buffer saline

6 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) -30.0 MIC 4 -30.0 40.030.0 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) -5.04.05.0 30.0/5.040.05.0 AD = aqua dest. PB = phosphate buffer, PBS = phosphate buffer saline

7 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 460 - BACTEC 460 Critical drug conc. (µg/ml) ---5.0 30.0/5.0-5.0 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 2.0 4.01.02.0 2.52.0 AD = aqua dest.

8 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) -0.52.0-8.00.5/4.0-0.25 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) -----1.0- AD = aqua dest. PB = phosphate buffer

9 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

10 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

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

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