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Bestimmung der Luftkeimzahl in Reinräumen mittels Fluoreszenzmarkierung

Katrin Husmann a , Bernd Wiedemann b und Klaus-Jürgen Steffens a

Pharmazeutische Technologie a und Pharmazeutische Mikrobiologie b
des Pharmazeutischen Institutes der Rheinischen Friedrich-Wilhelms-Universität Bonn

Detection of Airborne Viable Germs in Cleanrooms with a Fluorescence Marking Method

The monitoring of airborne viable germs in pharmaceutical cleanrooms with conventional methods reveals results only after 5 days. With the fluorescence method, a rapid and automatic technology, exists an alternative to conventional microbiological methods. The method offers the possibility to detect one germ per analysis. A new developed air sampler was used that samples microorganisms quickly and gentle into liquid media (1 m³ air with in 5-10 min). Comparison with 3 other commonly used air samplers shows similar results for all 4 samplers. To detect a small number of airborne viable germs with the fluorescence marking method (carried out with a ChemScan system) the results demonstrated good correlations to the conventional method, the plate count method. With the ChemScan system often more germs were detected than with the plate count. Furthermore, the fluorescence marking was more sensitive to some microorganisms (like Epicoccum nigrum). The air specimens which were taken with the new air sampler showed with the ChemScan system events which proved to be particles. The number of events that needed to be validated were less than 50, so that the validation with an epifluorescence microscope could be carried out in 5 min. The employment of the fluorescence marking method in combination with the new air sampler seems to be very successful for the aim of a rapid monitoring of airborne viable germs.

Key Words Fluoreszenzmarkierung · Keimzahlbestimmung · Luftkeimsammler · Reinräume