1.0 OBJECTIVE
To lay down a procedure is to provide personnel monitoring in the aseptic area.2.0 SCOPE
This procedure is applicable for personnel monitoring in all aseptic area.3.0 RESPONSIBILITY
Microbiologist4.0 ACCOUNTABILITY
Head QA & QC5.0 PROCEDURE
Microbiological Personnel monitoring is carried out as per following methods1. Finger Dab Method.
2. Gown swabbing at different places by two methods.
A. Surface Swab method
B. Contact Plate Method
5.1 Finger Dab Method
5.1.1 Prepare SCDA medium as per SOP for preparation of culture media.5.1.2 Aseptically pour approximately 15 – 20 ml of sterile molten cooled (40°C) SCDA agar into sterile 90 mm Petri plates.
5.1.3 Allow solidifying the plates under LAF, after solidification label all the plates with the name of media, preparation batch No. and date of preparation.
5.1.4 Invert and incubate the plates for preincubation to check the plates for any contamination if there is contamination discard the plates as per SOP for the destruction of microbial waste by Autoclaving.
5.1.5 Transfer the container in pass box of sterile area and personnel must be entered through Airlocks (Personnel entry) as per SOP for Entry and gowning procedure for the sterile area.
5.1.6 Collect the stainless steel container from pass box of sterile area and disinfect the container with sterile 70% IPA solution.
5.1.7 Label all the plates with the date of sampling, Personnel and hand name and Shift with the help of marker pen.
5.1.8 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 follow the same procedure.
5.1.9 Close the plate and immediately, disinfect the fingers with sterile 70% IPA solution and ask Operator to go to change room and replace with fresh sterile gloves and gown.
5.1.10 Prepare a positive control plate by streaking any pure culture of Salmonella / S. aureus / P. aeruginosa, on the surface of SCDA plate. For negative control, incubate the plate as it is without streaking.
Invert and incubate all the plates at 20 – 25°C for 72 hrs and 30 to 35°C for 48 hours.
5.1.11 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
5.2 By Surface Swab method
5.2.1 Prepare a normal saline solution by weighing accurately 0.9 gm of Sodium chloride AR/LR grade and dissolve in PW/ WFI. Finally, makeup volume to 100 ml with PW / WFI5.2.2 Take cotton Swab wet in normal saline solution to the tube of swab and Similarly prepare the required number of swab and place in a 1000 or 3000 ml beaker
5.2.3 Sterilize normal saline solution and prepared swab by Autoclaving at 121°C and 15-psi pressure for 20 minutes as per SOP for Sterilization by Autoclaving.
5.2.4 After Autoclaving swab to microbiology lab and allow to cool at room temperature
5.2.5 Transfer the swab to the aseptic area in an SS container through pass box of the material entry of sterile area and personnel must be entered through Airlocks (Personnel entry) as per SOP for Entry and gowning procedure for sterile area
5.2.6 Collect the material from pass box of the material entry of sterile area and disinfect with sterile 70% IPA solution.
5.2.7 Open the container, remove the swab from the tube and rub on the surface to cover approx 25 cm2 area, uniformly and horizontally in one.
5.2.8 Close the tubes and label with person name, location name, date. Carry out the same procedure for all locations and analyze the samples without store a long period.
5.2.9 Transfer the swab aseptically to MLT lab of microbiology lab through pass box and place all the sample swabs under LAF.
5.2.10 Unscrew the tubes and remove the swab stick by gently squeezing and then aseptically streak into a preincubated SCDA medium plate.
5.2.11 Label the preincubated SCDA medium Petri plate with person name, Date and location of swabbing.
5.2.12 Simultaneously prepare a positive control, by streaking bacterial culture suspension (Salmonella or S.aureus or P.aeruginosa) into a preincubated SCDA medium Petri plate.
5.2.13 Invert and incubate all the plates at 20 – 25°C for 72 hrs and 30 to 35°C for 48 hrs
5.2.14 After incubation, count the number of cfu formed on the plates with the help of colony counter and record the results cfu per 25 cm2.
5.3 By contact plate
5.3.1 Prepare the Contact media plates and pre-incubated the prepared plate as per media preparation SOP to check the contamination.5.3.2 Transfer the contact plates to the aseptic area in an SS container through passbox of the material entry of sterile area and personnel must be entered through Airlocks (Personnel entry) as per SOP for Entry and gowning procedure for the sterile area.
5.3.3 Aseptically open the Contact media plates & gently touch plates for surface sampling at selected places on the gown.
5.3.4 Label the contact SCDA medium plate with person name, Date and location.
5.3.5 Close the plate & keep it in an inverted position.
5.3.6 Clean the area of Gown by using disinfectant from where the sample is taken.
5.3.7 Remove the sampled gown to send for washing and autoclaving at provided change room. Wear another Gown before carrying out the work.
5.3.8 Invert and incubate all the plates at 20 – 25°C for 72 hrs and 30 to 35°C for 48 hrs. Record the observations.
5.4 Precaution
5.4.1 Result should be expressed as CFU per 25-cm2 in case of swab method.5.4.2 Maintain aseptic condition during testing.
5.4.3 Personnel monitoring shall be done in the airlock.
5.4.4 After finger dab testing, immediately sanitized the hands with sterile 70% IPA solution.
5.4.5 Result must be expressed per five fingers in case of finger dab method.
5.5 Frequency: Personnel monitoring in the sterile area - Daily after/during filling.
5.6 Limits/ Acceptance Criteria
Personnel Monitoring
Grade Gloves Personnel Gown
A <1 5
B 5 5
Finger Dab: Grade A <1cfu/plate, B < 05cfu/plate,
Gowns: < 05cfu/plate.
6.0 ABBREVIATIONS
SOP: Standard Operating ProcedureQA: Quality Assurance
QC: Quality Control
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