Rapid Detection Methods for Mycoplasma

 
 

Mycoplasma detection solutions that won’t keep you waiting.

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MilliPROBE® and MilliPrep™ Solutions for Culture-Free, Fast and Reliable Detection of Mycoplasmas

MilliporeSigma’s product range for real-time detection of mycoplasmas encompasses

  • MilliPROBE® Mycoplasma (based on TMA technology) and
  • MilliPrep™ sample preparation for Mycoplasma DNA and RNA

providing results comparable to those obtained with compendial mycoplasma testing but significantly faster - within a few hours or less!
These systems can be used to screen for Mycoplasma at various points in your process to

  • Strengthen your process risk mitigation plan
  • Keep your process moving forward feeling confident that your product is Mycoplasma free.

Mycoplasma - Real-Time TMA Technology for Fast, Sensitive, Reliable and Culture-Free Detection of Mycoplasmas

MilliPROBE®MilliporeSigma’s MilliPROBE® Mycoplasma system is an in-process early warning system that will provide you with faster, more effective detection of mycoplasmas in your bioreactor. The MilliPROBE® system combines speed, sensitivity and reliability in a single test system, enabling you to detect a Mycoplasma contamination event earlier in your process. It provides presence/absence results within four hours, compared to 28 days for traditional detection methods. This allows you to pinpoint contamination earlier, limiting the impact of the event and reducing downtime.

MilliPROBE® Mycoplasma: Robust and Reliable Detection in Only 4 Hours
Roka Bioscience
  • Completely closed assay design for protection from contamination and resulting false positives
  • Full system internal control 
  • Real-Time Transcription Mediated Amplification¹ (TMA) technology targeting abundant ribosomal RNA for increased sensitivity
  • Added sensitivity due to sample volumes of up to 20 mL
  • Designed to meet the European Pharmacopeia 2.6.7 guidelines
A Molecular Assay Anyone Can Run
  • Easy, closed, three step sample preparation
  • Semi-automated workflow requiring minimal hands-on time
Fast Implementation With Full Support and Validation Services
  • Feasibility and Method Development Services
  • Installation and On-site Training
  • Validation Protocol
  • On-Site IQ/OQ Validation Services
  • PQ Consulting Services

¹Real-Time Transcription-Mediated Amplification (TMA) is a technology of ROKA Bioscience

MilliPrep™ Sample Preparation Kit for Use with Nucleic Acid Based Rapid Detection Method for Mycoplasma

MilliPrep Sample Preparation Kit for MycoplasmaMilliporeSigma’s MilliPrep™ Sample Preparation Kit for Mycoplasma couples membrane based sample filtration and magnetic bead based nucleic acid purification for easy and representative sample preparation. The unique design of the MilliPrep™ device allows users to easily purify Mycoplasma DNA and RNA from their samples in three simple steps. Only minimal training is required to gain sample preparation proficiency. The closed design feature of the MilliPrep™ device also protects the sample from cross contamination, which can lead to false positive results and serious implications for your manufacturing processes. Unlike the many sample preparation methods that permit only small volumes, the filtration-based MilliPrep™ device can process up to 20 mL. A greater amount of target material leads to improved sensitivity.

MilliporeSigma’s MilliPrep™ Magnetic Bead Kit for Mycoplasma effectively purifies and concentrates the nucleic acids contained in the lysate recovered from the sample preparation device. This highly purified DNA and RNA sample is ready for use with any molecular rapid detection method.

By combining membrane-based filtration and magnetic bead nucleic acid purification, MilliPrep™ can effectively remove inhibitory substances which may interfere with the subsequent step of nucleic acid amplification.

MilliPrep™: An Easy to Implement Method that Enhances Productivity in Your Lab
  • Easy, closed, three step sample preparation
  • Streamlined workflow requiring minimal hands-on time
  • Prepares both DNA and RNA in the same sample for subsequent amplification and detection 
Filtration and Magnetic Bead Sample Preparation Method for Representative Results
  • Closed design for minimal risk of cross contamination
  • Sample volumes of up to 20 mL of bioreactor matrix for increased sensitivity
  • Membrane based sample preparation technique removes potential inhibitors of subsequent amplification step

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The MTC-NI System: Detection of Mycoplasmas Within 75 Minutes

The Mycoplasma Tissue Culture Non-Isotopic Rapid Detection (MTC-NI) Mycoplasma rapid detection system from MilliporeSigma: An easy-to-use first-line screening system for mycoplasmas:

  • Detection and analysis within 75 minutes (only 15 minutes of which are hands-on time)
  • Commonly occurring Mycoplasma species detected with a sensitivity of down to 105 microorganisms
  • Gen-Probe® Hybridization Protection Assay (HPA) technology for detection of multicopy rRNA, ensuring sensitivity and robustness
  • Highly tolerant to sample matrice variation and common inhibitors (Heparin, EDTA etc)
  • Ideal as a first-line screening tool for broad spectrum detection of mycoplasmas
  • Easy to use assay requiring only minimal training
How the MTC-NI System Works

MTC-NI SystemThe MTC-NI (Mycoplasma Tissue Culture Non-Isotopic Rapid Detection) system utilizes the patented HPA assay format from Gen-Probe: A labeled (acridinium ester) ssDNA probe which is complimentary to a conserved region of ribosomal Mycoplasma RNA is hybridized to the released rRNA of the target organisms. After the probe material binds to the target RNA, the acridinium ester is protected inside the newly formed double helix. When hybridization is completed (all DNA probes have found their target RNA), a selection reagent (0.6M sodium borate) is added to the solution. The selection reagent hydrolyses unbound probes, thus preventing any signal generation from non-hybridized probes. In the subsequent detection step, the bound probes will produce chemiluminescence (induced by hydrogen peroxide from the added detection reagent), which a Leader®  luminometer detects. It records the signal and displays it in Relative Light Units (RLUs), with a positive signal being defined as a value above a certain threshold.

The main advantage of this technology is the simplicity of the assay in terms of both handling and assay-components. Because the MTC-NI HPA assay uses a hybridization event followed by non-enzymatic hydrolysis and detection, it is highly tolerant to sample matrice variation and common inhibitors (Heparin, EDTA etc). These can have a strong negative affect on other widely used nucleic acid detection methods.

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