HIV Drug Resistance Training Module 13: Equipment and Supplies Topics Overview of Responsibilities Equipment and Supplies Needed for Genotyping Equipment Use and Care.
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HIV Drug Resistance Training Module 13: Equipment and Supplies 1 Topics Overview of Responsibilities Equipment and Supplies Needed for Genotyping Equipment Use and Care 2 Objectives Identify specialized equipment and supply needs for a genotyping lab. Describe how to ensure quality through appropriate care and use of equipment. 3 A Systems Approach to Laboratory Quality Organization Personnel Equipment Stock Management Quality Control Data Management SOP, Documents & Records Occurrence Management Assessment Process Improvement Specimen Management Safety & Waste Management 4 Functioning Equipment is Vital to Quality Service Produces reliable test results Functioning Equipment Lowers repair costs Prevents delays in testing Quality Results Maintains productivity 5 overview of responsibilities What are the responsibilities of lab management? What are the responsibilities of lab staff/technicians? 6 Management Responsibilities In the area of… Lab mgt. must ensure… Ordering pieces of equipment Each piece meets needs and specifications of lab Receiving and installing equipment The lab is ready for installation Accountability and supervision Assigning oversight responsibility for equipment Standard operating procedures Develop/delegate and approve equipment SOPs Operating equipment Train staff Troubleshooting Train staff Inventory control Establish inventory record Maintenance Develop maintenance program 7 Staff/Technician Responsibilities Attend training and learn all procedures Properly use the instrument by following SOPs Perform routine maintenance – Routine function check – Troubleshooting – Maintenance log Take corrective actions: notifying lab manager when problems arise Keep records on maintenance and problems 8 Discussion How will you assign responsibilities for equipment among different members of management and staff? 9 equipment and supplies needed for genotyping What equipment and supplies do all labs need for HIVDR testing? What equipment and supply decisions depend on the testing method? 10 Core Laboratory Equipment Microcentrifuge Vortex Biological safety cabinet Agarose gel electrophoresis Computer, printer, Internet connection UV Imaging system Reliable power supply (Uninterruptible power supply, UPS, for short outages) 11 Core Laboratory Supplies Micropipettes Agarose gel electrophoresis, power supply Refrigerators and freezers Distilled, deionized water supply Sink Ice machine Timers 12 Equipment for Genotyping May be influenced by platform (ViroSeq, TruGene, or Inhouse) DNA sequencer PCR machine Dead-air cabinet (PCR workstation) Computer attached to sequencer Specialized software dependent on Platform used Sequencing kit 13 Laboratory Supplies Sequencer supplies: capillary array, polymer, buffers PCR supplies: tubes, plates, trays RNA/DNA purification: columns, buffers Electrophoresis: Agarose, ethidium bromide or other stains, buffers, MW markers/standards Sterile, RNAse-free water Pipette tips (aerosol-resistant for pre-PCR) Polaroid film or special printing paper Disposable gloves Lab coats (disposable or not) 14 Types of Pipettes Single, multichannel, automated, repeater Use disposable, single-use, pipette tips Positive displacement and aerosol barrier tips recommended for pre-PCR work 15 Equipment: Pre-Amplification Area, In-house Assay Essential: – – – – – – – – Bench with sink / tap water Biohazard flow, class IIb Dead air cabinet (master mix preparation) -20°C freezer and 4°C refrigerator Microcentrifuge 12,500-15,000 g Vortex Timers Dedicated set of micropipettes for master mix and RNA extraction Optional: – Ultracentrifuge 21,000-25,000 g – Dead air cabinet (RNA extraction) – -80°C freezer 16 PCR Workstations (Dead-air Cabinets) 17 Equipment: Post-Amplification Area, In-house Assay Essential: – – – – – – – – – – – – – Bench with sink / tap water Dead air cabinet (nested reaction) Thermal cyclers (e.g. ABI, Bio-Rad, Hybaid, etc) Agarose gel apparatus Photo documentation of agarose gel DNA sequencer (e.g. ABI, Beckman) Computer + programs (editing) Microcentrifuge 450-550 g Vortex Dedicated set of micropipettes Timers -20°C freezer 4°C refrigerator Optional: – -80°C freezer 18 ABI Sequencing Instruments 19 Alternative Sequencing Instruments Beckman CEQ8000/8800 TruGene 20 Remember Workflow! Ice machine LAB 4 "Postamplification" Lab Sequencer NA extraction equipment Gel electrophoresis area PCR machines Eye wash workbench workbench workbench Centrifuge workbench "Specimen Processing" Lab sink Lab coat rack Lab coat rack PCR machines Eye wash Dead air cabinet Ice machine LAB 2 -20°C freezer PCR machines sink -20°C freezer LAB 3 Eye wash BSC LAB 1 Eye wash Lab coat rack -20°C freezer BSC -80°C freezer Lab coat rack sink Ice machine -80°C freezer NA extraction equipment -80°C freezer Lab coat rack workbench -80°C freezer Ice machine "Specimen Processing" Lab Centrifuge BSC Dead air cabinet -20°C freezer BSC LAB 2 Passthrough window "Nested PCR" Lab sink Ice machine LAB 1 "No Template" Lab Lab coat rack -20°C freezer -20°C freezer Centrifuge sink Centrifuge Consider lab layout (module 4) when planning equipment needs and placement workbench "Postamplification" Lab Sequencer Gel electrophoresis area PCR machines BSC: bio-safety cabinet 21 Reflection What is our “shopping list?” 22 equipment use and care What can we do to make sure our genotyping equipment is properly cared for? 23 Maintenance Properly maintained equipment is needed for quality results Maintenance: Routine and biannual or yearly scheduled maintenance – Reduces breakdowns – Reduces costs in the long run – Minimizes "down time" 24 Introducing New Equipment Calibrate or verify specified performance upon installation – – – – Pipettes Centrifuge Vortex Etc. Perform assay equivalence testing – Key components of the genotyping process (sequencer, PCR machine) – 10-20 samples previously reported and verify results within expected range of variability 25 Maintenance: Function Checks Verify that equipment is working properly Performed routinely – Daily, weekly, monthly – After adjustment or repair Example: – Monitoring refrigerator temperatures – Verifying pipette accuracy – Checking centrifuge speed 26 Refrigerator and Freezer: Use and Care Keep organized Periodically clean inside and outside Ensure door is completed sealed when closing CAUTION! – DO NOT store food items or beverages in laboratory refrigerator or freezer 27 Refrigerator & Freezer: Temperature Checks Monitor daily – Refrigerator: 4°C (2°C to 8°C) – Freezer: -20°C (+/- 5°C) or -80°C (+/- 10°C) If possible install electronic monitoring and alarm system Avoid "frost-free" freezers for storage of critical reagents (enzymes, specimens, etc.) 28 Refrigerator & Freezer: Temperature Log 29 Keep a Log for All Maintenance Activities 30 Exercise: Create a Maintenance Activity List Purpose: – Use what you have learned in this module and create a maintenance checklist specific to your test site Process: – Work in groups of 3-4 (or by test site) – Create a list of maintenance activities on a daily, weekly, monthly, and yearly basis – You can use the handout as a starting point. – Activity time = 10 minutes 31 Discussion What equipment and supplies do all labs need for HIVDR testing? What equipment and supply decisions depend on the testing method? What challenges do you face in your setting regarding equipment and inventory supply? 32 Summary Overview of Responsibilities Equipment and Supplies Needed for Genotyping Equipment Use and Care 33