Monitoring of Surface swabs in Non Aseptic processing area

1.0 OBJECTIVE

1.1 To describe the procedure for monitoring the surfaces of the non aseptic processing area by surface swabbing, sampling procedure, sampling locations, sampling time and frequency, incubation and interpretation of results.

2.0 SCOPE

2.1 This is applicable to surface swabs sampling in non aseptic processing area of Production block.

3.0 RESPONSIBILITY

3.1 Microbiologist

4.0 ACCOUNTABILITY

4.1 Head – Quality control

4.2  Head – Quality Assurance 

5.0  PROCEDURE                     

5.1  Requirements

5.1.1 Sterile swabs

5.1.2 Sterile water for injection for moistening swabs

5.1.3 90 mm petri plates with sterile soya bean casein digest agar media containing 0.5% glycerol v/v  and 4% Polysorbate 20 v/v.

5.2 Media and Plate preparation

5.2.1 Prepare the media as per SOP. and additionally incorporate 4 % polysorbate 20 v/v in the media meant for plates that are to be used for monitoring surfaces.

5.2.2 Prepare the plates as per SOP   

5.3 SWABS

5.3.2 Wrap the swabs with parchment paper in a clean and sanitized area.

5.3.3 The day before the swab sampling, pack the swabs in a beaker and autoclave the swabs and water collected in test tubes for moistening the swabs for sampling

5.4  PLATES

5.4.2 Use sterile plates prepared as in 5.2 to monitor the non aseptic processing area.

5.4.3 Collect the required number of plates from the incubator that are preincubated at least for 48 hours.

5.4.4 Inspect each plate thoroughly  for any colony, suspect colony, crack, condensate or dehydration in a black and white background.

5.4.5 If any such plate is observed record in the log book and discard as per SOP.

5.4.6 Transfer the inspected good plates in a clean sanitized canister and take to the IPQA pass box.                                                               

5.5 SAMPLING PROCEDURE

5.5.1 Follow the dress code and procedure for personnel entry to Helmut block and component preparation area.

5.5.2 Take the plates and sterilized swabs to the component preparation area on a trolley and sanitize the plates in the component preparation area.

5.5.3 Take the sterile swab, sterile water and the sanitized plate to the location where the sampling has to be done.

5.5.4 Take out the sterilized swab from the beaker and remove the parchment paper and hold the swab at one end.

5.5.5 Moisten the other end of the swab in sterile water.

5.5.6 Swab the sampling site by rolling the swab in 25cm2 area in a rolling fashion vertically and horizontally.

5.5.7 Streak the sampled swab on the petri dish containing sterile Soybean Casein Digest Agar with 0.5% glycerol v/v and 4 % polysorbate 20 v/v.

5.5.8 Similarly take the swabs at all the locations as given in the attached format Annexure I.

5.5.9 Moisten one swab with sterile water and streak directly on the agar surface to serve as Negative control.

5.5.10 Moisten one swab in a mixed culture of bacteria and fungi, and streak on the agar surface to serve as positive control. Record the no. of colonies observed.

5.5.11 Record the details of activity, name of the personnel, sampled by, time of sampling etc. in the format annexure II.

5.5.12 Collect all the plates and transfer the plates to the laboratory in the same canister used for bringing the plates.

5.6 INCUBATION

5.6.1 Transfer the plates to the walk-in incubator in QC lab at 30 –35°C.

5.6.2 Incubate the plates in inverted position for 48 hrs at 30 –35°C and read the plates for bacterial colonies after completion of 48 hrs.

5.6.3 If there is a holiday then read the plates on the next working day.

5.6.4 While reading, record the results for each plate in the format – Annexure I.

5.6.5 Transfer the plates to the other walk-in incubator having 20-25°C and incubate for next 72 hrs. Read the plates for mold and yeast colonies on each day of incubation since mold may overgrow the entire surface of the plate.

5.6.6 If there is a holiday then read the plates on the next working day.

5.6.7 While reading, record the results for each plate in the format – Annexure I.

5.6.8 After completion of incubation of the plates, take the plates having counts for identification of the isolates obtained.

5.6.9 Give the reference number to each isolate as per the SOP  and enter the reference number into the format – Annexure I.

5.6.10 Carry out the identification of isolates as per SOP .

5.6.11 After completion of the incubation period, fill format, sign, check and submit to QA.

5.7 Sampling frequency:

5.7.1 All locations as given in annexure I should be monitored once in a month.

5.8 Interpretation of Results:  

5.8.1 If count is found to be more than the specified alert limit, inform QA, write the corrective action report .

5.8.2 The occurrence must be investigated as per SOP  and remedial action taken to prevent or minimize its recurrence.

5.8.3 Appropriate action may include examination of Production records for any indication of an incident that would explain the finding, imposing additional monitoring and / or re-sanitizing the involved area with a suitable disinfectant. 

5.8.4 It may also be necessary until acceptable conditions have been restored.

5.8.5 The action taken should be fully documented in the standard format.

5.8.6 The technical staff to take corrective action should review the trend analysis of plate counts of both critical and non-critical areas frequently.

5.9 Acceptance criteria

Classified Area        Alert limits                         Action limits                                           

LAF                         NMT 5 cfu / 25cm2           NMT 10 CFU/ 25cm2          

6.0 ANNEXURES

Annexure – I Viable particulate monitoring record for Surface swabbing