1.0 Objective
To lay down the Standard Operating Procedure to provide guidelines for personnel monitoring in the sterile area2.0 Scope
This procedure is applicable to all sterile area at the manufacturing facility.3.0 Responsibilities
3.1 Implementation: Microbiologist3.2 Execution: Assistant Manager – Microbiology
4.0 Accountability
QC Manager
5.1.2 Clean SS container
5.2.2 Sterile 70% IPA solution
5.4.2 Aseptically pour approximately 15–20 ml of sterile molten cooled (45°C) SCDA agar into sterile 90 mm Petri plates.
5.0 Procedure
5.1 Equipment Required
5.1.1 Sterile dress5.1.2 Clean SS container
5.2 Material Required
5.2.1 Sterile SCDA plates5.2.2 Sterile 70% IPA solution
5.3 Utilities Required
5.3.1 NA5.4 Methodology
5.4.1 Prepare SCDA medium as per SOP for preparation of culture media.5.4.2 Aseptically pour approximately 15–20 ml of sterile molten cooled (45°C) SCDA agar into sterile 90 mm Petri plates.
5.4.3 Allow solidifying the plates at the room, after solidification label all the plates with the name of media, preparation batch No. and date of preparation.
5.4.4 Invert and incubate the plates at 30 to 35°C for 24 hrs. After incubation checks the plates for any contamination if there is contamination discard the plates as per the latest SOP for Destruction of Microbial waste by Autoclaving.
5.4.5 After pre-incubation, label all the plates with the date of sampling, Personnel name and Shift with the help of marker pen and wrap with aluminum foil and then keep in clean SS container
5.4.6 Transfer the container to the sterile area through pass box and personnel must be entered through Air locks as per latest SOP for Entry and gowning procedure for the sterile area.
5.4.7 Collect the materials from pass box and disinfect the container with sterile 70% IPA solution.
5.4.8 Call operator or personnel to be monitored, open the plate and tell personnel to place his/her right-hand fingers with gloves gently on the surface of SCDA plate. Use a fresh plate for left-hand finger and fallow the same procedure.
5.4.9 Immediately, disinfect the fingers with sterile 70% IPA solution and observe his/her finger carefully for the absence of media.
5.4.10 Close the plate and fallow the same procedure for all personnel.
5.4.11 Prepare a positive control plate by streaking any pure culture of E. coli / Salmonella abony/ Staph. aureus/ Ps. aeruginosa or wild culture, on the surface of SCDA plate. For negative control, incubate the plate as it is without streaking.
5.4.12 Invert and incubate all the plates at 20 – 25°C for 72 hrs and 30 to 35°C for 48 hours.
5.4.13 After incubation, count the number of CFU formed on the plates with the help of colony counter. Operate the colony counter as per SOP and record the results per 5 finger.
5.5.2 After finger dab testing, immediately wash the hands with sterile 70% IPA solution.
5.5.3 Result must be expressed per five fingers.
5.5.4 All pre-incubated plates should be rejected if a single plate shows evidence of microbial contamination.
5.7.2 Action Limit – 3 CFU/five fingers
5.4.4 Invert and incubate the plates at 30 to 35°C for 24 hrs. After incubation checks the plates for any contamination if there is contamination discard the plates as per the latest SOP for Destruction of Microbial waste by Autoclaving.
5.4.5 After pre-incubation, label all the plates with the date of sampling, Personnel name and Shift with the help of marker pen and wrap with aluminum foil and then keep in clean SS container
5.4.6 Transfer the container to the sterile area through pass box and personnel must be entered through Air locks as per latest SOP for Entry and gowning procedure for the sterile area.
5.4.7 Collect the materials from pass box and disinfect the container with sterile 70% IPA solution.
5.4.8 Call operator or personnel to be monitored, open the plate and tell personnel to place his/her right-hand fingers with gloves gently on the surface of SCDA plate. Use a fresh plate for left-hand finger and fallow the same procedure.
5.4.9 Immediately, disinfect the fingers with sterile 70% IPA solution and observe his/her finger carefully for the absence of media.
5.4.10 Close the plate and fallow the same procedure for all personnel.
5.4.11 Prepare a positive control plate by streaking any pure culture of E. coli / Salmonella abony/ Staph. aureus/ Ps. aeruginosa or wild culture, on the surface of SCDA plate. For negative control, incubate the plate as it is without streaking.
5.4.12 Invert and incubate all the plates at 20 – 25°C for 72 hrs and 30 to 35°C for 48 hours.
5.4.13 After incubation, count the number of CFU formed on the plates with the help of colony counter. Operate the colony counter as per SOP and record the results per 5 finger.
5.5 Precaution
5.5.1 Maintain aseptic condition during testing.5.5.2 After finger dab testing, immediately wash the hands with sterile 70% IPA solution.
5.5.3 Result must be expressed per five fingers.
5.5.4 All pre-incubated plates should be rejected if a single plate shows evidence of microbial contamination.
5.6 Frequencies
5.6.1 Shift wise5.7 Limits/ Acceptance Criteria
5.7.1 Alert limit – 1 CFU/five fingers5.7.2 Action Limit – 3 CFU/five fingers
6.0 Abbreviations
6.1 SOP: Standard Operating Procedure6.2 QA: Quality Assurance
6.3 QC: Quality Control
6.4 NA: Not Applicable
6.5 SCDA: Soybean Casein Digest Agar
6.6 IPA: Isopropyl Alcohol
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