Cambio - Excellence in Molecular Biology

Cell Culture Contamination Control

Cell Culture Contamination Control: Detection

Mycoplasma Detection

As an efficient contamination control measure we recommend frequent routine testing of cell culture materials with Venor®Gem Mycoplasma Detection Kits. These PCR- and qPCR-based mycoplasma assays allow fast (within a few hours), simple, effective and sensitive detection of mycoplasma-contaminated cell cultures in any standard molecular biology laboratory.

Regular mycoplasma testing and control procedures can significantly help in avoiding the costly consequences of contamination ensuring safe biopharmaceuticals and reliable scientific results.  Many scientific journals currently do not accept submissions of cell-based studies without evidence that mycoplasma testing has been performed.

To meet different customer needs Minerva Biolabs has developed a range of Venor®GeM mycoplasma detection assays using conventional PCR or qPCR methods which enable rapid and reliable identification of mycoplasma in cell cultures, culture supernatants and tissue culture media.

Conventional PCR

  • Venor®GeM Classic allows fast, reliable and time-saving routine monitoring of mycoplasma contamination in cell culture supernatants, media and biopharmaceuticals in research and industry by conventional PCR.

  • Venor®GeM OneStep is a kit for direct detection. It requires minimal pipetting, the kit includes all reagents required for PCR in a ready-to-use lyophilized reaction mix. Only samples or controls need to be added prior to PCR.

  • Venor®GeM Advance is designed for direct detection. Designed to reduce the total assay time and the pipetting steps, the kit contains PCR strips of tubes, each pre-dispensed with all reagents necessary for each PCR reaction, including the polymerase. For additional convenience, the gel loading buffer and dye are already included in the reaction buffer. After PCR, the products can be loaded directly on an agarose gel.

Real-time PCR (qPCR)

Our qPCR kits are TaqMan®-based qPCR Assays with labelled probes:  FAM® channel (mycoplasma) and HEX® channel (internal control to check for PCR inhibition from contaminants in sample).  So not only does the qPCR approach remove a lot of hassle, but the internal controls also give you reassurance that your negatives are not false.

  • Venor®GeM qEP utilises quantitative, real-time PCR (qPCR) for fast (approx. 3 h) and reliable screening of cell culture supernatants for mycoplasma contamination. The kit can be used in combination with any type of real-time PCR cycler able to detect the fluorescence dyes FAM® and HEX®. For clinical diagnostics in which human samples are tested we recommend using this kit as it conforms to EP 2.6.7, USP <71> and JP G3. It is suitable for both research and industry where certified compliance is needed or desirable. 

  • Venor®GeM qOneStep is a kit based on a real-time PCR (qPCR) assay for rapid, robust and sensitive detection of mycoplasma contamination. The detection procedure can be performed within 3 hours. Primers, nucleotides, probes, polymerase and an internal amplification control are provided as a ready-to-use, lyophilized reaction mix. This kit is a very straightforward and is highly suitable for academic labs and industrial applications where certified compliance of kits is not required.


Mycoplasma 10CFU™ Sensitivity Standards

10CFU™ Sensitivity Standards for validation of detection limit and robustness for individual sample material according to EP 2.6.7 with inactivated mycoplasma particles

Minerva Biolabs

Catalogue No.DescriptionPack SizePriceQty
102-0002Mycoplasma setall listed species £1,498.00 Quantity Add to Order
102-1003Mycoplasma arginini3 ampoulles 10CFU each £278.00 Quantity Add to Order
102-1103Mycoplasma salivarium3 ampoulles 10CFU each £278.00 Quantity Add to Order
102-2003Mycoplasma orale3 ampoulles 10CFU each £278.00 Quantity Add to Order
102-3003Mycoplasma gallisepticum3 ampoulles 10CFU each £278.00 Quantity Add to Order
102-4003Mycoplasma pneumoniae3 ampoulles 10CFU each £278.00 Quantity Add to Order
102-5003Mycoplasma synoviae3 ampoulles 10CFU each £278.00 Quantity Add to Order
102-6003Mycoplasma fermentans3 ampoulles 10CFU each £278.00 Quantity Add to Order
102-7003Mycoplasma hyorhinis3 ampoulles 10CFU each £278.00 Quantity Add to Order
102-8003Acholeplasma laidlawii3 ampoulles 10CFU each £278.00 Quantity Add to Order
102-9003Spiroplasma citri3 ampoulles 10CFU each £278.00 Quantity Add to Order

Description

10CFU™ Sensitivity Standards10CFU box

European Pharmacopoeia 2.6.7 Mycoplasmas requires a sensitivity of 10 CFU/ml sample volume for NAT-based methods like PCR to replace the traditional culture method. This feature of the test method must be shown by the performing lab as part of the robustness testing in presence of the sample matrix. As most cell culture labs and production facilities cannot accept vital mycoplasma in their facility or do not have access to a microbiology lab able to cultivate mycoplasma, these preparations allow safe and reliable validation of the procedure.
The mycoplasma have been cultivated in culture broth described in EP 2.6.7, titrated immediately in culture broth and plated for quantification in colony forming units (CFU/ml). Each dilution series has been performed in multiple by different operators for highest precision. The mycoplasma broth was harvested in the early logarithmic phase of the growth to avoid an atypical high ratio of dead mycoplasma particles and correspondingly a high GU*:CFU ratio. All strains have been obtained from the National Collection of Type Cultures (NCTC), UK, and cultivated in low passages.

Kit Components

3 vials with 10 CFU of the corresponding mycoplasma species, 2 negative control vials

For the mycoplasma set:
2 vials with 10 CFU of each mycoplasma species listed in the EP 2.6.7 (18 vials in total), 2 negative controls

Result evaluation

Use of a suitable Mycoplasma PCR kit, e.g. Intego® Mycoplasma

Required Consumables

None

Required lab devices

Regular lab equipment (centrifuge, pipettes, etc.)

Shelf Life and Storage

Kit components are maintainable at 2 to 8 °C for at least 12 months.

10CFU graph

Fig. 10 CFU have been resuspended in cell culture medium, 200 µl DNA extracted with the MB DNA Extraction and mycoplasmal DNA amplified with Intego® Mycoplasma. PCR was performed on a Corbett RotorGene®6000.

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Protocols

Click here for protocols

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Applications & Benefits

For validating robustness and sensitivity of NAT-based mycoplasma test methods in presence of the sample matrix.

Each vial contains 10 CFU of inactivated mycoplasma. By adding the sample matrix of interest a sample according to EP 2.6.7 is prepared which has to be tested positive by the method applied. Obviously, the inactivated sample material is not suitable for the culture method anymore. As a result of proficiency tests on NAT-methods for mycoplasma detection it became obvious that in means of highest sensitivity DNA extraction is indispensible. The extract can directly be used for PCR.

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Related products

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