QIAcuity Mycoplasma Quant Kit and Standards

For digital PCR quantification and detection of mycoplasma contamination using the QIAcuity Digital PCR System

S_1345_3_LS_dPCR_QIAcuity_Mycoplasma_Quant_kit

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QIAcuity Mycoplasma Quant Kit

Cat. No. / ID:  250261

OneStep Advanced Probe Master Mix, OneStep Advanced RT Mix, OneStep Enhancer GC, QIAcuity Mycoplasma Assay, QIAcuity Mycoplasma Internal Control, QIAcuity Mycoplasma Positive Control, RNase-Free Water
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$2,000.00
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KitStandard
QIAcuity Mycoplasma Quant Kit
QIAcuity Mycoplasma Standard CFU Kits
The QIAcuity Mycoplasma Quant Kit and Standards is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

  • Probe-based assay to quantify and detect 127 different mycoplasma species
  • Pharmacopeia-compliant workflow, including 10 different Mycoplasma CFU Standards  for validation and for optional positive sensitivity controls.
  • Third-party validation report available

Product Details

The QIAcuity Mycoplasma Quant Kit reliably detects and quantifies the presence of mycoplasma rRNA in various sample types. Combined with a recommended sample prep method, we offer a NAT (Nucleic Acid Technique) workflow for mycoplasma testing that is compliant with the European, US and Japanese Pharmacopeia. The robust process eliminates time-consuming cultivation of mycoplasma. The workflow is particularly suited to sample types of varying purity (for example, cell bank samples, in-process samples, such as virus harvest and final lots/batches).


The QIAcuity Mycoplasma Quant Kit provides a consistent, accurate, precise and highly reproducible detection of mycoplasma presence. The detection via rRNA enables a higher sensitivity than using DNA because of the multiple copies present in a single bacterial cell. The assay is still able to detect DNA if the RT-step is skipped, enabling a high degree of flexibility.

The kit is compatible with the QIAcuity Digital PCR System and the QIAcuity Nanoplates.

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Performance

Use the QIAcuity Mycoplasma Quant Kit to detect mycoplasma contamination in cell cultures and pharmaceutical products during the different manufacturing steps. Benefit from a workflow that is fully compliant with the European, US and Japanese Pharmacopeia. Download a full validation report to assess the performance of the kit and its standards. Please note, you must verify the validation for your specific sample matrix before first use.

 

Principle

The QIAcuity Mycoplasma Quant Kit, together with the QIAcuity Digital PCR System, form a unique solution for detecting and quantifying mycoplasma contamination. The digital PCR-based workflow offers a winning combination of performance, ease of use and compliance with the European Pharmacopeia (EP; chapter 2.6.7), the US Pharmacopeia (UP; chapter <63>), and the Japanese Pharmacopeia (JP; chapter G3-14-170). Besides the 10 mycoplasma species mentioned in the Pharmacopeias, you can detect another 127 mycoplasma species using the QIAcuity Mycoplasma Quant Kit. To assess the performance of the workflow, download a validation report from an independent party, Minerva Biolabs GmbH (Berlin, Germany).    


The validated workflow consists of only two steps. First, extract the nucleic acids from the sample, then analyze the nucleic acids via RT-dPCR. With the QIAcuity OneStep Advanced Probe chemistry, you can perform the reverse transcription and PCR in a single step. Although we recommend detecting mycoplasma using an RT-dPCR protocol, you can also directly detect DNA in a digital PCR reaction without the RT-step.

The principle of the dPCR reaction in the nanoplates is described here.

 

Procedure

The validated workflow consists of two steps in which the nucleic acids are extracted from the sample and subsequently analyzed via RT-dPCR. Using the QIAcuity OneStep Advanced Probe chemistry, the required reverse transcription and PCR are done within a single hands-on step. Although we recommend detecting mycoplasma using the RT-dPCR protocol described below, the assay also detects DNA and can be used in a dPCR reaction without the RT-step.  


Sample preparation    


To reliably detect mycoplasma contamination, RT- and PCR inhibitors and other impurities should be absent in your sample. As an overall control for inhibitors, you can choose to add the QIAcuity Mycoplasma Internal Control to the lysis buffer before the sample extraction. You can directly use the eluate for mycoplasma detection using the QIAcuity Mycoplasma Quant Kit with the corresponding QIAcuity OneStep Advanced Probe chemistry.


QIAcuity OneStep Advanced Probe Kit


Merge reverse transcription and PCR for detecting mycoplasma contamination without setting up a separate RT-reaction by combining HotStart Reverse Transcription (RT) Enyzme, HotStart QuantiNova DNA Polymerase, proprietary chemical components and the QIAcuity Mycoplasma Assay. 


QIAcuity Mycoplasma Assay    


The QIAcuity Mycoplasma Assay includes a FAM-labeled 20x primer-probe mix for the detection of a specific mycoplasma rRNA target and a HEX-labeled primer-probe mix for the optional detection of the QIAcuity Mycoplasma Internal Control. We’ve thoroughly tested this duplex reaction for cross-talk. With the assay, you can rely on a single control for RNA extraction, reverse transcription, amplification and detection, eliminating the need for extra reactions.

 

 

Applications

QIAcuity Mycoplasma Quant Kit and Standards are designed for detecting and quantifying mycoplasma contamination in cell cultures and biopharmaceutical development processes.

Resources

Quick-Start Protocols (1)
Kit Handbooks (1)
Safety Data Sheets (1)
Certificates of Analysis (1)

FAQ

How long can I store the reconstituted Internal Control and the reconstituted Positive Control?
Reconstituted QIAcuity Mycoplasma Internal Control aliquots can be stored at 30°C to 15°C for up to six weeks. More than five freezethaw cycles should be avoided.
FAQ ID - 3997
Do you recommend storage of highly diluted mycoplasma samples?
No, it is not recommended to store mycoplasma eluates in a highly diluted state.
FAQ ID - 3998
Can I use Internal Control as overall control?
Yes! You can spike-in the QIAcuity Mycoplasma Internal Control to the lysis buffer and proceed with the sample isolation as usual. The Internal Control will then be detected by a second assay in the yellow channel of the QIAcuity Digital PCR System and you can calculate the recovery efficiency.
FAQ ID - 3996
Is the kit compatible with unpurified in-process samples?
Technically yes, but we recommend performing sample purification following the handbook.
FAQ ID - 3994
My samples are expected to have low concentrations. Can I load more sample into the workflow or more lysate into the PCR reaction?
The amount of sample tested for the sample preparation is 250 µL. This is the best condition for both samples containing a high or low amount of background cells. Depending on your sample, you can optimize the amount of sample to isolate. You can load a maximum of 22.6 µL eluate per RT-dPCR reaction. Due to the high sensitivity of the workflow, this should be enough to get a signal for very low concentrations of mycoplasma contamination as described in the different Pharmacopoeias.
FAQ ID - 3995