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Fluorescence-based Rapid Microbial Detection Tec

Fluorescent-based detection methods allow for faster microbial quality control throughout the manufacturing process. The ability to obtain microbial tests earlier enables one to keep pace with increasing demand and allows corrective actions in case of product contamination. The Milliflex® Quantum system is an easy-to-use, non-destructive, fluorescent staining-based system ensuring consistent and accurate microbiological results while reducing overall time to result.

Fluorescent Staining For Rapid Microbial Detection

The Milliflex® Quantum system is based on membrane filtration and fluorescent staining for rapid detection of microorganisms. After filtration and incubation, the reagent is applied to the membrane to stain any viable and culturable microorganisms with a fluorescent marker. The reaction requires active microbial metabolism for enzymatic cleavage of a non-fluorescent substrate. Once cleaved inside the cell, the substrate liberates free fluorochrome into the microorganism cytoplasm (Figure 1). As fluorochrome accumulates inside the cells, the signal is naturally amplified. The cells are then exposed to the excitation wavelength of the fluorescent dye so that they can be visually counted.

Fluorescent staining mechanism for rapid microbial detection

Figure 1.Principle of Fluorescent Staining.

Milliflex® Quantum Rapid Detection System

The Milliflex® Quantum system is designed for the rapid detection and enumeration of microorganisms over a broad range of filterable samples. The system detects microbial colonies even before they are visible to the naked eye. Milliflex® Quantum system combines widely used membrane filtration with proven enzymatic fluorescent staining of viable microorganisms for early detection. This significantly reduces the time to result compared to the traditional methods. The fluorescence-based quantitative detection method is fast, reliable, and robust, over the whole range of assays following the US and Ph. Eur. guidelines.

Milliflex® Quantum uses a non-destructive procedure, making it possible to identify any viable microorganisms subsequently using your current identification methodology.

Milliflex® Quantum system for rapid detection of microorganisms
  • Detection times for most microorganisms are between 8 – 48 hours
  • Compatible with any ID technology
  • Easy to use with a simple workflow, requiring minimal training
  • Results comparable to a compendial method, facilitating Milliflex® Quantum validation
  • Compact hardware with a small footprint
  • Economical and robust system

Milliflex® Quantum System Applications

Milliflex® Quantum system uses industry-standard membrane filtration techniques to detect viable and culturable microorganisms down to 1 CFU per sample. This easy-to-use and simple system is compatible with a wide range of filterable samples:

  • Raw materials such as media, buffers, pharmaceutical ingredients, and water
  • In-process samples (bioburden before sterilization) including CIP/SIP samples, media for fermentation, buffers for manufacturing, and intermediate process samples
  • Final product
  • Environmental samples

Milliflex® Quantum Testing Workflow

The Milliflex® Quantum system employs 3 easy steps for rapid bioburden testing using two proven technologies: membrane filtration and fluorescent staining. The system consists of a reader, camera, and fluorescent reagents used in combination with our Milliflex Oasis® system.

1. Sample Preparation

The Milliflex® Quantum method utilizes a device for sample preparation, which ensures consistent and reliable results. Filter the desired sample volume through the ready-to-use Milliflex Oasis® filtration device. Place the membrane filter base onto a prefilled agar cassette and incubate.

2. Fluorescent Staining

Transfer the membrane filter to a pad soaked with the fluorescent reagent and incubate for 30 minutes.

3. Counting CFUs

Count fluorescent colonies through the window of the Milliflex® Quantum reader or use the camera to view the colonies on your computer screen.

Agar plate after fluorescent staining for rapid microbial detection with the Milliflex® Quantum system

Visualize plate after staining. View of the membrane in the reader.

Agar plate, microbial membrane Alt text: Agar plate with membrane, no bacteria colonies visible

CFUs are not visible outside of the reader.

Reincubated agar plate with visibly grown bacteria colonies

After reincubation, CFUs are visible to the eye.

For reference, the tested sample of in-process, non-sterile water using the Milliflex® Quantum system is shown here. After detection, the membrane was reincubated for full growth and Identification.

4. Reincubation

Reincubate for identification of the microorganisms. Place the membrane filter on a prefilled agar media cassette for reincubation. Collect and isolate the microorganisms and identify them using any existing ID methodology.

Milliflex® Quantum Spot Counter Software

The Milliflex® Quantum camera combined with the Milliflex® Quantum Spot Counter software makes the sample reading more convenient and allows you to track records for optimized microbiology QC labs. Download the software here.

Validated by Industry Peers

Dr. Marcel Goverde

Dr. Marcel Goverde
Managing Director, MGP Consulting, Switzerland
“Milliflex® Quantum was successfully validated as a rapid alternative method to the traditional bioburden testing”

Dr. Tim Sandle

Dr. Tim Sandle
Ph.D., Head of Microbiology, Risk Management, and Sterility Assurance at Bio Products Laboratory Limited.
Read the study by Tim Sandle
Watch the on-demand webinar

Milliflex® Quantum Services

The Milliflex® Quantum platform is simple and economical, enabling the user to focus resources on the validation method. We also offer a comprehensive range of services to support the implementation, including IQ, OQ, and PQ guidelines. Our highly experienced team of experts is available to assist you. They can help you ensure that all test criteria lead to a completely qualified and accurate test method, including hardware, expendables, and product testing method (S.O.P.).

Our validation services include:

  • Validation strategy & PQ consultancy
  • On-site data analysis support and report generation
  • On-site support for the implementation of qualification tests
  • Operator technical training
  • Development of testing methods

Discover method development & validation services

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