Presentation on theme: "ASEPTIC AWARENESS Training Objectives: To further increase awareness about the microbial contaminants found on our bodies and in the environment. To discuss."— Presentation transcript:
ASEPTIC AWARENESS Training Objectives: To further increase awareness about the microbial contaminants found on our bodies and in the environment. To discuss behaviors to minimize contamination by personnel. To perform lab exercises to reinforce the concepts learned in class.
ASEPTIC AWARENESS Training What is Aseptic Processing? What processes are most vulnerable to contamination?
ASEPTIC AWARENESS Training Types of Contamination
ASEPTIC AWARENESS Training What are SPORES Why are they a A MAJOR CHALLENGE!!!! http://www.samedanltd.com/members/archives/PMPS/Spring2003/graphics/f1_p12.gif http://micro.med.harvard.edu/faculty/rudner.html
ASEPTIC AWARENESS Training Sources of Contamination
ASEPTIC AWARENESS Training CONTROLLING contamination Facility design Personnel AND visitors wear gowns appropriately Personnel AND visitors behave appropriately
ASEPTIC AWARENESS Training Facility Design: Clean Area Classification
ASEPTIC AWARENESS Training Facilities: General Cleanroom Design HEPA/ULPA filters on ceiling Exhaust vents on floor Drains in aseptic processing areas are inappropriate Airlocks and interlocking doors to control air balance Seamless and rounded floor to wall junctions Readily accessible corners Floors, walls, and ceilings constructed of smooth hard surfaces that can be easily cleaned Limited equipment, fixtures and personnel Layout of equipment to optimize comfort and movement of operators
ASEPTIC AWARENESS Training Facilities: HEPA Filters http://people.deas.harvard.edu/~jones/lab_arch/nano_facilities/hepa.gif High Efficiency Particulate Air Minimum particle collection efficiency: 99.97% for 0.3µm diameter particles. Disposable Filter made of pleated borosilicate glass microfiber
ASEPTIC AWARENESS Training Class 10,000 cleanroom http://www.americancleanrooms.com/am/photogallery_08.html Class 100 cleanroom Facilities: Cleanroom Classification
ASEPTIC AWARENESS Training Facilities: Cleanroom Classification
ASEPTIC AWARENESS Training Facilities: Pressure Differentials Used to maintain airflow in the direction of higher cleanliness to adjacent less clean areas A minimum of 10-15 Pascals should be maintained between the aseptic area and an adjacent rooms with differing cleanroom classifications (doors open)
ASEPTIC AWARENESS Training http://news.thomasnet.com/images/large/451/451402.jpg Facilities: Air Lock Permits the passage of objects and people into a cleanroom. Consists of two airtight doors in series which do not open simultaneously.airtightdoors Spray down materials with 70% IPA before placing in the airlock
ASEPTIC AWARENESS Training Fiber-shedding materials such as cardboard and paper –Cardboard packaging must be removed and items placed into non-cardboard containers. Wood (i.e. wooden pallets) Undesignated charts Facilities: Material NOT permitted in a Cleanroom
ASEPTIC AWARENESS Training Facilities: Cleaning Water should be changed FREQUENTLY 1. Vacuum all accessible surfaces 3. Mop floors using a lint free polyester mops attached to stainless steel handles 2. Wipe surfaces with a cleaning solution
ASEPTIC AWARENESS Training http://www.rit.edu/ http://www.imi.org.uk/ Avoid Cleanrooms When Sick!!!
ASEPTIC AWARENESS Training Avoid cleanrooms when ill Frequent bathing and shampooing Avoid getting sunburned Avoid cosmetics such as face powder, hair sprays, perfumes and aftershave Clothing should be clean, nonfrayed and nonlinting Avoid smoking Personnel: Hygiene
ASEPTIC AWARENESS Training http://www.engenderhealth.org/IP/surgical/sum4.html Personnel: Gloves When are gloves worn? What compromises gloves? How often should gloves be examined and sanitized?
ASEPTIC AWARENESS Training Qualifying Personnel Assess after gowning/gloving Microbiological surface sampling of several locations –Glove fingers –Facemask –Forearm –Chest –Periodic requalification is necessary
ASEPTIC AWARENESS Training Biological Safety Cabinets Small benchtop sized class 100 clean area
ASEPTIC AWARENESS Training Vertical Laminar Flow Area http://www.terrauniversal.com/products/hoods/vlfstation.php
ASEPTIC AWARENESS Training ISOPROPYL ALCOHOL Powerful disinfectant and antiseptic Mode of action: denatures proteins, dissolves lipids and can lead to cell membrane disintegration Effectively kills bacteria and fungi
ASEPTIC AWARENESS Training Particles >= 0.3µm emitted per minute Personnel Activity Snap Smock Membrane Coverall No Movement100,00010 Light Movement500,00050 Heavy Movement1,000,000100 Change Position2,500,000250 Slow Walk5,000,000500 Personnel: Behaviour Austin Contamination IndexSource: Encyclopedia of Clean Rooms, Bio-Cleanrooms and Aseptic Areas, Dr. Philip Austin, PE, 2000 Minimize movement: Work slowly and purposefully Note: Light/heavy movement refer to partial body movements (motioning with arm, tapping toes, etc.). Change of position refers to whole body motion (standing up, sitting down, etc.).
ASEPTIC AWARENESS Training ENVIRONMENTAL MONITORING In aseptic processing, one of the most important laboratory controls is the environmental monitoring program Guidance for Industry: Sterile Drug Products Produced by Aseptic Processing -Current Good Manufacturing Practice, FDA, September 2004
ASEPTIC AWARENESS Training Microbial Monitoring Methods Touch or Contact plates –RODAC Plates Swabs www.esa.int Environmental Monitoring: Surface Monitoring http://www.blood.co.uk/hospitals/services/Micro/Bact2.htm
ASEPTIC AWARENESS Training Environmental Monitoring: Particulate Air Monitoring
ASEPTIC AWARENESS Training Active Air Monitoring RCS Plus Passive Air Monitoring Settling Plates RCS: Rotor Centrifugal Sampler Environmental Monitoring: Viable Microbial Air Monitoring
ASEPTIC AWARENESS Training Environmental Monitoring: Trending Data Why is it important? Evaluate the disinfection efficiency Are certain microbes migrating into the aseptic processing area from a lesser controller area? Long and short term Data by: –Shift –Room –Shift –Operator –Product –Container type –Filling line –Sampling –Testing personnel
ASEPTIC AWARENESS Training http://www.arches.uga.edu/~kristenc/cellwall.html QC Micro: Identifying Microbes
ASEPTIC AWARENESS Training Gram Stain QC Micro: Identifying Microbes
ASEPTIC AWARENESS Training Metabolic Based Assays Vitek QC Micro: Identifying Microbes
ASEPTIC AWARENESS Training QC Micro: Identifying Microbes Metabolic Based Assays
ASEPTIC AWARENESS Training Staphylococcus xylosus Reduction of Tetrazolium Violet QC Micro: Identifying Microbes Metabolic Based Assays
ASEPTIC AWARENESS Training QC Micro: Identifying Microbes Metabolic Based Assays
ASEPTIC AWARENESS Training QC Micro: Identifying Microbes Genotype Based Assay: MicroSeq
ASEPTIC AWARENESS Training QC Micro: Identifying Microbes Genotype Based Assay: MicroSeq PCR: Polymerase Chain Reaction
ASEPTIC AWARENESS Training SEQUENCE THE DATA QC Micro: Identifying Microbes Genotype Based Assay: MicroSeq
ASEPTIC AWARENESS Training ENDOTOXIN LIMIT FOR WFI IS 0.25EU/ml QC Micro: LAL Assay
ASEPTIC AWARENESS Training The Lysate QC Micro: LAL Assay
ASEPTIC AWARENESS Training Turbidimetric - increase in turbidity. Chromogenic - addition of a peptide that when cleaved during the assay turns a distinct color. Endpoint - Measure turbidity/absorbance after a definitive time period. Kinetic - Measure rate of increased turbidity/absorbance. Gel Clot – a clot forms and stays intact at the bottom of the assay tube when it is inverted QC Micro: LAL Assay TYPES of LAL ASSAYS
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