Application Note

Rapid Microbial Detection Of Contaminants In Water Samples

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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 can 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.

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 labeling of viable and culturable micro-organisms collected on a Milliflex® mixed cellulose esters (MCE) membrane. After filtration and incubation, micro-organisms retained on the membrane are labeled 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 reincubation after the staining step, allowing for further identification using existing ID methodologies.

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