1.0 OBJECTIVE :
- To provide a procedure for Microbial Evaluation by Swab Method.
2.0 SCOPE :
- This procedure applies to the microbial evaluation of bioburden and pathogen on various the surface like wall, floor, tank, drainage point, equipments parts etc. The data shall be collected for monitoring purpose.
3.0 RESPONSIBILITY :
- Microbiologist
4.0 ACCOUNTABILITY:
- Head of Department
5.0 PROCEDURE :
5.1 PREPARATION OF COTTON SWABS :
- Preparation of dry swabs: Swabs are prepared on clean sticks using clean cotton. These swabs are then placed in glass Petri plates or glass test tubes and sterilized by autoclaved at 121°C for 15 minutes. The sterile swabs can be used within 24 hours.
- Alternatively, use ready available sterile swabs as bought out item with certificate.
- Sterile Saline: Prepare physiological saline ( 0.90 % NaCl in water ), sterilize at 121 °C for 15 lbs for 15 minute. Use for moistening the swab before swabbing.
5.2 REQUIREMENTS:
- Sterile forceps
- Sterile Hand gloves
- 70% IPA
- Sterile Petri dishes
- Sterile cotton swabs
- Sterile test tubes
- Sterile Tips
5.3 Procedure for Swab test:
- Moisten a sterile dry cotton swab in sterile saline. Swab approximately 25 cm2 for surface area as per requirement and for drain use dry swab to take the sample from drain. Transfer these swabs to 10 ml sterile saline.
- Preserve the sample in refrigerator, if the sample is not analysed same day.
- Transfer 1 ml saline sample in sterile two Petri plate add 15 to 20 ml SCDA media for bacterial count and SCA media for fungal count respectively allow to solidify the media.
- Incubate the plates for 5 days, SCDA at 30°C to 35°C for bacterial counts and SCA 20°C to 25°C for fungal counts and record result in cfu/25cm2 for drain swab sample, for Equipment part and surface and Tank cleaning in cfu/25cm2
- Transfer reaming saline sample in 90 ml SCDM and mix well.
- Incubate SCDM at 30°C to 35°C for 24 hours and analyse the pathogens, as per the SOP.
- Reclean area as per SOP in case the limit crosses.
- Record the observation in Annexure No.: 01.
5.4 FREQUENCY:
- Applicable For Drain, Equipments – 6 Months or as per requirement ±15 days.
- Tank cleaning : 3 Months or as per requirement ±15 days.
5.5 LIMITS / ACCEPTANCE CRITERIA:
- For Drain Sample:
- A) Bacterial count :
| Alert limit | Action limit | Maximum limit |
| 60 cfu/25cm2 | 80 cfu/25cm2 | 100 cfu/25cm2 |
- B) Fungal count : Absent
- C) Pathogens : Absent
- For Equipment Parts and Surface:
- A) Bacterial count :
| Alert limit | Action limit | Maximum limit |
| 60 cfu/25cm2 | 80 cfu/25cm2 | 100 cfu/25cm2 |
- B) Fungal count : Absent
- C) Pathogens : Absent
- For Tank:
- A) Bacterial count :
| Alert limit | Action limit | Maximum limit |
| 60 cfu/25cm2 | 80 cfu/25cm2 | 100 cfu/25cm2 |
- B) Fungal count : Absent
- C) Pathogens : Absent
6.0 ABBREVIATIONS:
| Abbreviation | Expanded form |
| SCDM | Soyabean Casein Digest Medium |
| SCDA | Soyabean Casein Digest Agar |
| SCA | Sabouraud Chloramphenicol Agar |
| SOP | Standard Operating Procedure |
| °C | Degree Celsius |
| ml | Milliliter |
| cfu | Colony Forming Unit |
| cm2 | Centimeter squire |
| IPA | Iso Propyl Alcohol |
7.0 ANNEXURES:
| Annex. No. | Title |
| 01 | Microbial Evaluation By Swab Method |
8.0 SOP REFERENCES
- GMP Guidelines
END OF THE SOP
ANNEXURES :
Annex. No. 01 Microbial Evaluation By Swab Method
