FRESH PERSPECTIVES
Milliflex® Quantum – Detection of Microbial Contaminants in Water Samples
Beth Brescia EMD Millipore Corporation
Water is a key raw material utilized in the manufacturing of products within the pharmaceutical, biopharmaceutical and healthcare industries. Within each industry, regulatory requirements exist for microbial contamination in different levels of water purity. The microorganisms found in these water systems are mainly stressed, slow-growing strains characterized by long incubation times (5 to 7 days) before growth can be detected using traditional microbiology methods. The time required before contamination can be detected in water may cause delays in product release, and extend the storage time of products. By using rapid detection methods, manufacturers are able to address contamination events sooner, avoid line shutdowns, release product to the market faster, and reduce warehousing costs [1].
The Milliflex® Quantum system is a convenient platform for the rapid detection of microbial contamination in filterable samples. The system is based on a non-destructive fluorescence labelling of viable and culturable microorganisms collected on a Milliflex® mixed cellulose esters (MCE) membrane. After filtration and incubation, microorganisms retained on the membrane are labelled by a fluorescent marker. The reaction consists of an enzymatic cleavage by active microorganism metabolism of a non-fluorescent substrate. Once cleaved inside the cell, the free fluorochrome is liberated into the microorganism cytoplasm. As fluorochrome accumulates inside the cells, the signal is naturally amplified. The cells are then exposed to the excitation wavelength of the fluorescent dye in the Milliflex® Quantum reader so that the colonies can be counted visually. Detected micro- colonies can be recovered by membrane re-incubation after the staining step allowing for further identification using existing ID methodologies [2].
Materials • Milliflex® Funnels, Mixed Cellulose Esters (MCE) 0.45µm (MXHAWG124)
• Milliflex® R2A Cassettes (MXSMCRA48) • Milliflex® Liquid Cassette (MXLMC0120) • Milliflex® Quantum Reagent Kit (MXQTV0KT1)
Beth Brescia is an Application Scientist at EMD Millipore Corporation. She has worked in the
Pharmaceutical and Biotechnology industries for over 20 years. Beth received her Bachelor’s degree in Medical Technology and her Master’s degree in Laboratory Science from Quinnipiac College and is certified as a Six Sigma Black
Belt. Since 2011, Beth has been at EMD Millipore Corporation working on Rapid Microbiology
methods in the Applications Laboratory within BioMonitoring. Corresponding author – email:
beth.brescia@
emdmillipore.com.
Equipment • Milliflex® PLUS Pump (MXPPLUS03)
• Milliflex® Quantum System (MXQUANK01): reader, camera, stand, membrane transfer tool, removal rack, and Quantum Spot Counter software
• Incubator, 32.5 +/- 2.5°C Method Overview
Various water samples (lab water and purified water) were tested in order to assess the ability of the Milliflex® Quantum System to detect microbial contaminants quicker and to evaluate the reduction of time-to-final result in comparison with the compendial method.
The water sample was filtered through a Milliflex® 0.45 µm MCE membrane using the Milliflex® PLUS pump system. The membrane was placed onto a prefilled R2A Milliflex® agar media cassette
50 American Pharmaceutical Review | Fresh Perspectives 2013
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