Fundamental Autoclave Techniques

Transcription

1 Fundamental Autoclave Techniques Version 3, September 2010 Veronika Tatarinoff GSBmE University of NSW Brandy Nelson University of Kentucky

2 The following information is directed at aseptic/sterile surgical, tissue culture (OGTR/PC2) and QC1 techniques at the GSBME.

3 GSBME Autoclave Training Program This presentation forms part of the GSBME Autoclave training program. 1. Fundamental Autoclave Techniques (FAT) 2. Review of supporting literature (SOP, SWP, RA) 3. FAT quiz 4. Demonstration of GSBME Autoclave use 5. Competency assessment 6. Authorisation sign off in WITA

4 Learning outcomes : At the end of this session participants will be able to: Describe the mechanisms of autoclave sterilisation Identify autoclave/s at GSBME Identify materials that can be autoclaved Package and label items for sterilization Correctly load/unload the autoclave Identify and select appropriate sterilisation cycle/s Verify autoclave sterilisation process Maintain log of sterilisation Dispose of biowaste appropriately Identify and use the Personal Protective Equipment (PPE) and devices in a safe manner. Identify hazards and risks to self and others, improper autoclave techniques and identify appropriate responses to injury.

5 What does aseptic mean? preventing putrefaction [rotting, decomposition] by securing the absence of bacteria Oxford Dictionary Disinfection. The process of destroying or inhibiting the growth of micro-organisms on a surface. Sterilisation. The destruction of all living micro-organisms, including viruses and spores. This is an absolute state.

6 Micro-organisms Include bacteria, viruses, fungi & spores Found on every external surface including respiratory tract and alimentary canal A micro-organism infection occurs when the micro-organism grows in an area where it isn t normally found or in numbers greater than normal. Surgical and cell culture procedures provide an ideal opportunity for micro-organisms to cause problems.

7 Sterilisation methods There are a number of methods which can kill or remove micro-organisms including: Heat (dry or steam autoclave) Gas (eg Ethylene Oxide) UV or gamma irradiation H plasma Liquid sterilisation (eg glutaraldehyde) Filter sterilisation (liquids only)

8 Autoclave use basics Fundamental autoclave information Personal Protective Equipment (PPE) Packaging Loading / Unloading Operating cycles User logs Hazards Improper Autoclave Practices

9 Critical Sterilisation parameters Autoclaving uses high temperature, high pressure and steam to kill organisms. The critical autoclaving parameters are: temperature steam under pressure time Items have to be hot enough for long enough in the presence of steam to disrupt the proteins of the micro-organisms.

10 Autoclave (steam) sterilisation Water is boiled under pressure and the steam is enclosed within the chamber. Objects placed in the chamber and the steam circulates around and penetrates them. Temperatures of C at set pressure for minutes are generally used. Condensation creates negative pressure and draws in additional steam. Moist heat kills microorganisms via coagulation of proteins.

11 What can be autoclaved? Surgical Instruments Glassware Plastic tubes and pipette tips Solutions and water Animal food and bedding Waste

12 What can t be autoclaved? The effectiveness of autoclaving depends on steam, the steam must be able to penetrate the material being autoclaved. Goods that are packed in impervious packaging cannot be autoclaved. Because damp heat is used, heat labile products (such as some plastics) cannot be sterilised in this way or they will melt. Items that may be damaged by steam, should be sterilised by another method. Chemicals (e.g. bleach) emit toxic fumes when autoclaved Incompatible chemicals e.g. acids, bases, organics solvents It is essential to ensure that your material/s can be autoclaved.

13 PPE for Autoclave Users Eye Protection Lab Coat, Buttoned Closed-toed Shoes Heat-resistant Gloves

14 Packaging for Autoclaving Be sure the material can be autoclaved No volatile chemicals, acids, bases, sharps Utilize containers and autoclave bags appropriate for autoclaving Clear or yellow biohazard bags Autoclave paper bags/roll Metal bins or containers Pyrex or Shott glassware Plasticware Do not overfill containers Do not tighten lids Label Identify contents, owner & date

15 Loading the Autoclave Load material to allow efficient steam penetration Autoclave clean items and waste separately Do not allow material being autoclaved to touch the sides or top of the chamber Ensure load probe cord is not in trolley path to prevent damage

16 GSBME AUTOCLAVE USE Consult with the Animal House manager if animal surgery is in progress before using the autoclave. Surgery has priority. ALWAYS CHECK Sterilisation parameter SETTINGS BEFORE USE. Different materials or quantities may require different sterilisation temperatures and times. There is a guide on the wall opposite the autoclave and in SOP S006. Specific instruction on operation of each GSBME autoclave is given in SWPI005 & I006 and will be demonstrated to you.

17 GSBME Autoclaves Getinge HS55 (automated) Atherton Benchtop model (small manual)

18 Getinge HS 55 Automated This Autoclave is a large stand alone model. It is controlled by a microprocessor and has both set programs and manual parameter selection. If your material requires specific parameters please see autoclave custodian.

19 P02 - POROUS CYCLE USE: DRAPES / WRAPPED PACKAGES / BOTTLES Items that are required to be dry at the completion of the cycle and/or requiring greater steam penetration. PARAMETER SETTINGS STERILISE 121 C 16 minutes DRYING TIME 5 minutes DRYING AIR PULSE 10 minutes

20 P09 - HARD GOODS CYCLE USE: INSTRUMENTS/SOLID ITEMS Items of equipment that do not require wrapping, drying or prolonged cooling. PARAMETER SETTINGS STERILISE DRYING TIME 121 C 16 minutes 5 minutes

21 P04 FLASH (HARD GOODS) CYCLE USE: INSTRUMENTS/SOLID ITEMS Items of equipment for surgical procedures that do not require wrapping, drying or prolonged cooling. PARAMETER SETTINGS STERILISE DRYING TIME 126 C 12 minutes 3 minutes

22 Liquids Liquids take longer to heat up and cool down than most solid materials. A control (similar fluid) at the maximum volume of sample load is required for autoclaving liquids. The load probe is placed in the control bottle to ensure that the liquids reach the correct temperature for the correct time to ensure sterilisation. Lids must be loose to prevent explosion. Liquids must be in heat treated laboratory glassware with room for fluid expansion. A liquid cycle must be used to prevent boiling over.

23 LIQUID CYCLE P05 Fo Liquids, open USE: SALINE, WATER Non heat sensitive liquids that require a probe in a control volume to ensure specified parameters are reached. A slow pressure release is used to avoid boiling over and allow cool down PARAMETER SETTINGS STERILISE DRYING RAMP TIME 121 C 16 minutes 0.05/m Ensure that lid is loose on the bottle/s during sterilisation

24 P10 - WASTE CYCLE USE: OGTR/PC2 TISSUE CULTURE WASTE BINS Items in the bins are contaminated with biological materials and do not require wrapping, drying or prolonged cooling. PARAMETER SETTINGS STERILISE 121 C 60 minutes DRYING TIME 5 minutes Following completion & verification of sterilisation, waste must be double bagged, labeled and placed into yellow bins in 4 C cold room pending disposal by UNSW contractor.

25 Bench top Autoclave The GSBME small benchtop model is a totally manual, gravity displacement style of autoclave with no drying or cool down cycle. Modification of parameter settings is made by adjusting gauges directly on the autoclave. Primarily this autoclave is a back-up if required.

26 Autoclave User Log User Log must be completed with every use User Logs are maintained for at least five years Date Time Authorised User Cycle Type e.g. fluid, hard goods Items /comments eg drapes, bottles, waste, validation

27 Unloading the Autoclave Put on Personal Protective Equipment Verify cycle conditions were met Stand clear of door when opening Allow contents to cool before removal Carefully remove items Be especially careful with fluids and plastic bins

28 How can you tell if something has been sterilised? INDICATORS can be used to ensure an autoclave has reached the correct temperature and pressure for the correct amount of time. They fall into 3 broad categories: * Chemical * Biological * Physical

29 Indicators Chemical indicators, can be found on medical packaging and autoclave tape, and these change colour once the correct temperature conditions have been met. This indicates that the object inside the package, or under the tape, has been exposed to heat. Physical indicators often consist of an alloy designed to melt only after being subjected to 121 C for 15 minutes. If the metal has melted, the change will be visible. Biological indicators include Attest devices. These contain spores of a heat resistant bacterium. If the autoclave did not reach the right temperature, the spores will germinate, and their metabolism will change the colour of a ph-sensitive chemical. Printout & gauges. As well as these separate indicators, autoclaves have temperature and pressure gauges visible from the outside. A printout maybe available to confirm conditions were met.

30 Example of a Chemical indicator Autoclave tape - colour change(bottom) indicates a temp of 80 C has been reached.

31 Examples of Physical indicators Untreated physical indicators Class 5 & 6 Colour change in treated (bottom) indicator, sterilisation confirmed

32 Examples of Biological indicators Unprocessed Processed - Failed Processed - Passed

33 Autoclave Cycle Verification Each Autoclave Load (automatic documentation) Have the sterlisation conditions been met? Document all autoclave runs in the User Log Review the printout report & any other indicator that you may have used. If conditions were met, remove the load. Printout report can be filed in lab book. For validation cycles indicator and print our are filed in the equipment validation log. If conditions were not met, do not remove the load, review and repeat cycle. If conditions are not met again, contact Autoclave custodian.

34 Packaging OGTR/PC 2 Waste for Autoclaving Tie waste bag with string, leave bag in metal biohazard bin Fix autoclave tape to bag Leave bin lid off to one side for autoclaving It is essential that each biowaste load has been verified as sterile prior to transfer to cold room pending removal by UNSW waste contractor. Indicator in the centre of the load

35 OGTR/PC2 Biowaste Bins Over full bin

36 Hazards associated with using autoclaves Burns from hot autoclave, sterilised items, steam. Sharp/hot/heavy items falling trolley onto operator. Cuts from broken glass Possible noxious/toxic fumes released from melted plastics Biohazard contamination (OGTR/PC2 waste) Chemical incompatibility In an emergency call x56666 security.

37 Importance of Proper Autoclave Use Tremendous pressure from steam in chamber provides explosive potential High temperatures and presence of extremely hot water creates potential for burns and scalding Inadequate decontamination allows for the potential of biological hazards and personnel and environmental contamination

38 Autoclave Explosion

39 Results of Improper Autoclave Use

40 Improper Autoclave Practices

41 Additional Information It is essential that you read and understand the following GSBME documents prior to your FAT quiz. 1. SOP S006 - Guideline to sterilisation at GSBME 2. SWPI005- Operation of getinge autoclave SWPI006- Operation of small autoclave 3. Risk Assessment - Basic use of Autoclaves It is essential that you have completed PC2 training, quiz and authorisation prior to entry to PC labs at GSBME. It is essential that you have completed UNSW Animal Care and Ethics training and have ACE approval to conduct any animal work.

42 Where do I go from here? This fundamental autoclave techniques is followed by a quiz. Download from OHS page on GSBME School Website: Submit your completed quiz to either the Lab or Animal Research Facility Manager who will arrange: Demonstration of autoclave/s use You will then complete a competency assessment Authorisation for autoclave use occurs when Signed off as competent by the trainer.

43 Don t forget your PPE!