CARIBBEAN REGIONAL MICROBIOLOGY STANDARD OPERATING PROCEDURES Media Preparation and Quality Control – SOP No: CRM-SOP 18 CAREC PAHO WHO EUROPEAN UNION STRENGTHENING OF MEDICAL LABORATORY SERVICES IN THE CARIBBEAN A CARIFORUM Project Funded by the European Union and Implemented by CAREC Media Preparation and Quality Control Title: Media Preparation and Quality Control SOP Number: CRM-SOP: 18 Version: 1 Page Number: 1 of 102 Prepared By: Caribbean Regional Standard Effective Date: 1st September 2007 Methods Drafting Group Review Date: 1st September 2008 TABLE OF CONTENTS Amendment procedure 4 Introduction 5 Media production process flow chart 5 1. Media Preparation and Quality Control 6 1.1 Introduction 6 1.2 Types of Media 6 1.3 Media Production 10 2.0 Media Quality Control Methods 11 2.1 Introduction 11 2.2 Principles of QC of Media 11 2.3 Shelf-life 11 2.4 Standardization of Laboratory Media 12 2.5 Control Medium 12 2.6 Test Programme 12 2.7 Physical Characteristics 12 2.8 Quality documentation 12 2.9 Methods 12 2.10 Examination of Physical Characteristics 13 2.11 Test for Microbiological Performance 14 2.12 QC Tests for Solid Media 14 2.13 QC Tests for Liquid Media 15 2.14 Interpretation of Microbiological Tests on Liquid Media 15 2.15 Sterility Tests 15 2.16 Quality Control Limits for Haemophilus Test Medium Agar 15 2.17 Storage of Culture Media 16 3.0 Quality Control of Culture Media Prepared In-house 16 3.1 Procedure 16 3.2 Quality Control Protocols 16 4.0 Quality Control of Commercially Prepared Media 18 4.1 Exempt Media Category 18 4.2 Nonexempt Media Category. 18 4.3 General QC Requirements for both Exempt and Nonexempt Media 19 4.4 Q.C. Requirements for Nonexempt Media Only 19 4.5 Reporting Deficiencies Identified by the User 19 5.0 Quality Control of Media not included in Section 6 20 6.0 Methods 24 6.1 Preparation of Alkaline Peptone Water 24 6.2 Preparation of Blood Agar 26 6.3 Preparation of Bile Esculin Agar 28 6.4 Preparation of Baird Parker Agar 30 6.5 Preparation of Bile Esculin Azide Agar 31 6.6 Preparation of Chocolate Agar 34 6.7 Preparation of CLED Agar 36 STRENGTHENING OF MEDICAL LABORATORY SERVICES IN THE CARIBBEAN 1 A CARIFORUM Project Funded by the European Union and Implemented by CAREC Media Preparation and Quality Control Title: Media Preparation and Quality Control SOP Number: CRM-SOP: 18 Version: 1 Page Number: 2 of 102 Prepared By: Caribbean Regional Standard Effective Date: 1st September 2007 Methods Drafting Group Review Date: 1st September 2008 6.8 Preparation of Cooked Meat Medium 38 6.9 Preparation of Brain Heart Infusion Broth 40 6.10 Preparation of Brain Heart Infusion Agar 42 6.11 Preparation of GN Broth 44 6.12 Preparation of Haemophilus Test Medium (HTM) Agar 46 6.13 Preparation of MacConkey Agar 48 6.14 Preparation of MacConkey Agar with Sorbitol 50 6.15 Preparation of Mueller Hinton (MH) Agar 52 6.16 Preparation of Nutrient Agar 54 6.17 Preparation of New York City (NYC) Medium 56 6.18 Preparation of Peptone Water 58 6.19 Preparation of Phenylethyl Alcohol (PEA) Agar 60 6.20 Preparation of Phenylalanine Agar 62 6.21 Preparation of Mannitol Salt Agar (MSA) 64 6.22 Preparation of Sabouraud Dextrose Agar 66 6.23 Preparation of Thayer Martin (TM) Agar 68 6.24 Preparation of Xylose Lysine Deoxycholate (XLD) Agar 70 6.25 Preparation of Thiosulphate Citrate Bile Salt Sucrose (TCBS) Agar 72 6.26 Preparation of Trichosel Broth 74 6.27 Preparation of Tetrathionate Broth 76 6.28 Preparation of Candida Bromcresol Green (BCG) Agar 78 6.29 Preparation of CHROMagar 80 6.30 Preparation of Lim Broth 82 6.31 Preparation of Kligler Iron Agar (KIA) 84 6.32 Preparation of Urea 86 6.33 Preparation of Simmons Citrate 88 6.34 Preparation of MILS Medium 90 6.35 Preparation of Triple Sugar Iron (TSI) Agar 93 7.0 References 96 Appendix 1: Preparation of Media (Flowchart) 97 Appendix 2: Preparation of Suspensions for Quality Control of Nonexempt Media 98 Appendix 3: Media Quality Control Record Sheet 99 Appendix 4: Media Quality Control Sheet 100 Appendix 5: Exempt and Non-exempt Categories for Media 102 STRENGTHENING OF MEDICAL LABORATORY SERVICES IN THE CARIBBEAN 2 A CARIFORUM Project Funded by the European Union and Implemented by CAREC Media Preparation and Quality Control Title: Media Preparation and Quality Control SOP Number: CRM-SOP: 18 Version: 1 Page Number: 3 of 102 Prepared By: Caribbean Regional Standard Effective Date: 1st September 2007 Methods Drafting Group Review Date: 1st September 2008 Acknowledgements The ‘Strengthening of Medical Laboratory Services in the Caribbean’ (SMLS) Project, would like to thank the following people for their participation in researching and writing the microbiology standard methods, and also the individual countries for supporting this initiative. Many thanks also for the facilitators of the standard methods meetings, the method editors and administrative support. Participants Country Name Organisation Aruba Ms. Astrid Dirksz Landslaboratorium Anguilla Ms. Everette Duncan Princess Alexandra Hospital, Stoney Ground Bahamas Mr. Allison Scavella Princess Margaret Hospital Barbados Ms. Gail Trotman Public Health Laboratory Ms. Juliana Applewaite Queen Elizabeth Hospital Mr. Edmund Blades Public Health Laboratory Belize Ms. Solitaire Parra Maza Central Medical Laboratory British Virgin Islands Ms. Allene Brewley Peebles Hospital Ms. June Greene Medicure Ltd. Ms. Wayveney Armstrong MEDICAL Diagnostic Laboratory Cayman Islands Mr. Dale Andrew Chin Caymans Islands Health Services Authority Curacao Mr. Osric Wanga Analytical Diagnostic Centre Ms. Helga Leito Analytical Diagnostic Centre Grenada Ms. Sonia Ann Edwards St Georges Hospital Guyana Ms. Alexis Wilson Pearson Georgetown Public Hospital Ms. Ede Tyrell Langevine University of Guyana Jamaica Ms. Rayaad Khan Central Medical Laboratories Ltd Ms. Valerie Levy UWI Microbiology Department Ms. Heather Wint Caledonia Medical Laboratory (Biomedical) Ms. Adriene Kellier Cornwall Regional Hospital Mr. Norman S. Burke National Public Health Laboratory St Lucia Mr. Martin S. Mc Kenzie Ezra Long Laboratory St Vincent and Mr. Elliot Samuel Milton Cato Memorial Hospital the Grenadines Caribbean Epidemiology Ms. Radha Gosein CAREC Centre (CAREC) Ms. Michele Nurse-Lucas CAREC Ms. Denise Clarke CAREC Dr. Ashok Rattan CAREC Trinidad Ms. Zobida Khan Mohammed Trinidad Public Health Laboratory (TPHL) Mr. Anthony Bayley Trinidad Public Health Laboratory (TPHL) Mr. Jawaheerlal Mewahlal College Of Science, Technology And Applied Arts of Trinidad and Tobago (COSTAATT) Ms. Monica Pollard Port of Spain General Hospital Dr. William Swanston Eric Williams Sciences Medical Complex Turks and Caicos Islands Ms. Lessonjule Lyons Doney Grand Turk General Hospital Laboratory Ms. Peggy Hermitt Myrtle Rigby Hospital STRENGTHENING OF MEDICAL LABORATORY SERVICES IN THE CARIBBEAN 3 A CARIFORUM Project Funded by the European Union and Implemented by CAREC Media Preparation and Quality Control Title: Media Preparation and Quality Control SOP Number: CRM-SOP: 18 Version: 1 Page Number: 4 of 102 Prepared By: Caribbean Regional Standard Effective Date: 1st September 2007 Methods Drafting Group Review Date: 1st September 2008 Facilitators/Editors Country Name Organisation Canada Dr Harold Richardson Quality Management Program – Laboratory Section Trinidad Ms. Julie Sims Strengthening of Medical Laboratory Services in the Caribbean (SMLS) United Kingdom Ms. Jacki Watts Bristol Health Protection Agency United Kingdom Ms. Valerie Bevan Health Protection Agency (HPA UK)* Administrative Support Function Name Organisation Administrative Support Ms. Stacey-Mae Stephens SMLS Project Administrative Support Ms. Reesa Moonsie SMLS Project Rapporteur Ms. Margaret Hunte Independent Graphic Design and Support Function Name Country Graphic Design Ms. Cathleen Jones Trinidad & Tobago Graphic Support Mr. Adrian Nicholls United Kingdom Graphic Support Ms. Karen Lara-Augustine Trinidad & Tobago *The Health Protection Agency, England, is acknowledged for the knowledge and expertise of their staff and working groups in the UK who develop National Standard Methods, on which the Caribbean Regional Microbiology Standard Operating Procedures are based. STRENGTHENING OF MEDICAL LABORATORY SERVICES IN THE CARIBBEAN 4 A CARIFORUM Project Funded by the European Union and Implemented by CAREC Media Preparation and Quality Control Title: Media Preparation and Quality Control SOP Number: CRM-SOP: 18 Version: 1 Page Number: 5 of 102 Prepared By: Caribbean Regional Standard Effective Date: 1st September 2007 Methods Drafting Group Review Date: 1st September 2008 INTRODUCTION The Caribbean Regional Standard Methods include a selection will be based on a review of EQA results, most variety of standard, validated methods, produced as common and/or critical tests. Part of the method a single standard operating procedure (SOP) for use in a standardization process will be an ongoing review and variety of levels of microbiology laboratory service. amendment procedure. The CSMDG consists of It is intended that these methods provide detailed microbiology laboratory representatives from most of instructions for microbiology services for microbiological the CARIFORUM countries, all of whom were nominated investigations, in order to provide accurate, reliable and to the task by the CRMC. reproducible results which will have clinical utility. These methods may be adopted by laboratories within the This initiative should enable the region to implement a region, or adapted, provided that such adaptations