To lay down the Procedure to test the efficacy of UV lamp against microbial challenge.
1.0 Purpose :
To lay down the Procedure to test the efficacy of UV lamp against microbial challenge.
2.0Scope :
This procedure applies to UV lamps of Pass box, Laminar Air Flow bench and Garments box located in the microbiology area.
3.0 Responsibility:
3.1 The responsibility of testing the efficacy of UV lamp lies with the Microbiologist/Analyst.
4.0 Definitions/Abbreviations: Nil.
5.0 Procedure :
5.1 Take the viable culture suspension of S.aureus/Ready made culture suspension(with traceable certificates) containing10-100 CFU.
5.2 Pipette 1 mL/ as per requirement of culture suspension into each of seven sterile petri dishes.
5.3 Add to each plate 15-20 ml sterile Soyabean Casein Digest agar that previously has been melted and cooled to approximately 45°C.
5.4 Cover the petri dishes, mix the culture suspension with the agar by tilting or rotating the dishes, and allow the contents to solidify at room temperature.
5.5 Expose the six agar plates contains culture suspensions by removing lid under the UV lamp of LAF Bench and one agar plate contains culture suspensions without exposing to UV lamp which act as positive control.
5.6 Prepare one agar plate with out adding culture suspension and this will act as negative control.
5.7 In every 5 minutes interval remove one exposed agar plate and label 1st plate as 5 minutes, 2nd plate as 10 minutes, 3rd plate as 15 minutes , 4th plate as 20 minutes, 5th plate as 25minutes and 6th plate as 30 minutes
5.8 After completion of exposure, close the plates. Invert the plates and incubate along with negative control and positive control at 30–35 °C for 48 hours.
5.9 The same procedure shall be followed for three consecutive days to validate the process.
5.10 The same above procedure shall be followed for UV lamps of pass box, garment box.
5.11 Observe and record the number of CFU in the Formatxxxx.
5.12 UV lamps shall be ON for 24 hours in pass box and garment box.
5.13 While transferring any material through UV pass box, material should be exposed under UV. UV exposure time should be equal to UV efficacy time.
5.14 Culture suspensions, samples, positive cultures are transferring through pass box into testing room should not be exposed under UV.
5.15 Once in 6 months/ as per requirement UV lamps shall be replaced and recorded in formatxxxx.
5.16 Acceptance criteria:
5.16.1 When there are no colonies observed at a specific duration of exposure time, the same timings shall be considered as UV efficacy time (exposure time of all UV lamps except the UV lamp of LAF).
5.17 Frequency: Once in three months ± 7 days.
6.0 Related Documents:
6.1. Procedure for sub culturing and preparation of viable culture suspension from pure cultures
6.2. Procedure for Operation and Usage of UV Pass Box.
Microbiology, microbial challenge, uv lamp
