Evaluation of Coriolis® microbial air sampler coupled with RMM
Alternative solution for rapid airborne contamination control
Results

Abstract
For environmental contamination control in cleanrooms, Bertin Technologies (France) has developed a technology dedicated to the monitoring ofairborne particles. The goal is to propose a sampling method compatible with Rapid Microbiological Methods (RMM) in order to get reliable & specificdata and tackle the impact on pharmaceutical production of time-to results within impaction method.
This microbial air sampler has been validated according to ISO14698-1 in terms of physical and biological efficiencies (HPA study – July 2008) : theresults show the Coriolis® µ is as efficient as the traditional method and even better for high particles diameter (results available on www.coriolisairsampler.com).
AES Chemunex has also validate a protocol coupling Coriolis® µ with ScanRDI® (cytometry) and allows to get the results in only 3 hours (fromsampling step to results). This study aims at completing this data and at testing different RMMs on the samples collected with Coriolis® µ.

Experiment A = comparison of Coriolis® µ and impactor (table 1)- Different areas controlled, equipments placed side by side, 2 replicates- Filtration of the whole sample from Coriolis® sampling (0,45µm filter) + filter placed on TSA agar plate- Incubation of plates 20-25°C for 72-96h + 30-35°C for 48-72h

Experiment B = Coriolis® µ sampling and ScanRDI detection (table 2)- Each liquid sample from Coriolis® splitted in A&B aliquots
A = filter plate method
B = filtered and analysed with ScanRDI

Table 1 - the impactor and Coriolis® µ giveequivalent results on the air samples in the lab.

Context
The microbiological quality control of environments aims at ensuring the quality of products in case of cleanrooms production /ensuring health and safety of workers and exposed people.
Accurate & reliable measurements of microbial contamination depend on the choice of an adapted air sampler / a representative sample fromthe controlled environment / the limitation of losses due to a failure of the sampler to capture particles containing micro-organisms or to due toinactivation of viable micro-organisms during collection so that formation of visible colonies on agar surfaces will not occur.

Objective : realize the feasibility study of the Coriolis® µ air sampler coupled with RMM (Scan RDI) inorder to implement it as a new solution for rapid investigation in case of contamination and forroutine monitoring in production sites.

Samples

Impaction

Coriolis®
+ filter plate

Table 2 - ScanRDI analysis gives higher counts than the filterplate method does, especially for contaminated air samples.
Samples

Coriolis® + Scan RDI

Coriolis®
+filter plate0Biohood

01M

Biohood

0Biohood00

Biohood+hands

10Counter top 26 (4 M +2 B)

2BCounter top 3

92 (1 M + 1 B)

4(2 M + 2 B)

Counter top + walking

66

7 (3 M + 4 B)

Counter top 21M

Counter top + hands

103

8B

Counter top 36 (3 M + 3B)

8 (7 M + 1 B)

Counter top 3

2B

6 (3 M + 3 B)

These higher results are partly due to the viable but non-culturable(VNC) microbials present in the air which can not be detected withtraditional impaction method allthough they can be pathogen.

M = Mold – B = Bacteria

Discussion

Material & methods
The air sampling has been carried out in one of the Micro Lab using either traditional air sampler (impactor) or Coriolis® µ. Samples
collected with the Coriolis® µ were further processed by filter-plate or ScanRDI method (RMM).

Liquid sample  Access to alternative methods  Rapid results(RMM beyond cultivable flora)

AIR SAMPLERS = impactor vs Coriolis® µ (Bertin Technologies)
The Coriolis®µ air sampler is based on a patented cyclonic technology: it concentrates airborne particles
into a liquid collection media.

The Coriolis® system is an easy to use portable air sampler that collects air samples into liquid media. This allows quick microbial
detection when coupled with RMM technologies as demonstrated into this study.
Coriolis® µ, Bertin Technologies

ANALYSIS METHOD = ScanRDI (AES Chemunex)
The ScanRDI® is based on cytometry and uses a double discrimination key : viability and cell membrane integrity.
Viability substrate
Enzyme

The feasibility study conducted in Saint-Louis Micro Lab indicates that Coriolis® system collects airborne microorganisms incomparable amount to the impaction system does. ScanRDI can be used as Rapid Microbiology Method coupled with Coriolis®
collection in order to give rapid results (around 3 hours from sampling to result). It is also shown here that the amount ofmicroorganisms detected with the ScanRDI is higher than with the traditional method as far as it is not based on cultivability but on
viability; appropriate alert and action limits may thus need to be re-evaluated for ScanRDI results. This couple of innovative methods
fits for the Environmental Monitoring application and could be implemented for investigation and routine monitoring in production
sites and in critical areas and cleanroom environments.

Microorganism
to be detected

Free fluorochrome

Acknowledgement
Key words : biocontamination – airborne particles – microbial airsampler – cleanrooms – alternative methods (RMM)• Pfizer study Team: Dr Lin Chen (responsible of the study), S. Fennell, W. Snapp• Bertin Technologies: Alexandra Guerin, Quitterie Desjonquères• AES Chemunex: Pierre Barbez