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.


MycoSPY® qPCR Master Mix

Real-Time Mycoplasma PCR test kit for rapid, highly sensitive detection of mycoplasmas in cell cultures

Biontex

Catalogue No.DescriptionPack SizePriceQty
M010-025MycoSPY® qPCR Master Mix25 Assays £183.00 Quantity Add to Order
M010-050MycoSPY® qPCR Master Mix50 Assays £315.00 Quantity Add to Order

Description

  • Detects all cell-culture-relevant mycoplasma strains
  • Highly sensitive
  • Master Mix with FAM/HEX fluorescence probes allows for easiest protocol (see workflow)
  • Fast results in less than 1.5 hours without the need for gel electrophoresis

A number of options are available for conclusive detection of the presence of mycoplasmas in contaminated cell cultures. A commonly but unreliable method is the DAPI staining of cells followed by microscopic analysis. However, PCR based methods give the most reliable results for routine checks. Biontex MycoSPY®, MycoSPY® Master Mix and MycoSPY® qPCR Master Mix detection kits are rapid, reliable and highly sensitive methods of PCR-based mycoplasma detection in cell cultures.

The MycoSPY® qPCR Master Mix consists of all necessary components for PCR analysis (hot-start Taq polymerase, buffer, dNTPs, primer, FAM/HEX fluorescence probes). The kit includes an Internal Control and PCR-grade water for rehydrating the lyophilized master mix. With a real-time PCR thermocycler suitable for detecting FAM and HEX fluorescent dyes, mycoplasma can be detected quickly and easily in cell cultures.

 

If mycoplasmas have been detected in your cell culture, the highly effective antibiotic mixture MycoRAZOR® eliminates the contamination rapidly and efficiently.

 

 

Workflow

 

The Master Mix with integrated FAM/HEX fluorescence probes is designed for maximum ease of pipetting and subsequent detection in real time:

Workflow for MycoSpy qPCR Master Mix

 

 

More than a quarter of all animal cell cultures are contaminated with mycoplasmas / mollicutes. The strains most commonly found in cell cultures, with a total probability of 94%, are: M. fermentans (47%), M. hyorhinis (19%), M. orale (10%), M. arginini (9%), A. laidlawii (6%) and M. hominis (3%). In addition, the following strains were found with lower probability: M. gallisepticum, M. pneumoniae, M. salivarium, M. synoviae and S. citri. All these mollicute strains, in addition to a variety of other strains, are detected by MycoSPY® qPCR Master Mix. BLAST analysis was used to verify of the primer specificity. The list shows only Mollicute strains with 100% probe and primer match.

 

List of mollicute strains detected with MycoSPY® qPCR Master Mix

Mycoplasmas
M. agalactiae M. crocodyli M. lagogenitalium M. pulmonis
M. alligatoris M. cynos M. microti M. salivarium
M. alvi M. dispar M. moatsii M. sualvi
M. amphoriforme M. edwardii M. molare M. synoviae
M. arginini M. felis M. mucosicanis M. testudineum
M. bovigenitalium M. fermentans M. muris M. testudinis
M. bovis M. gallisepticum M. mustelae M. verecundum
M. buccale M. genitalium M. mycoides M. volis
M. canadense M. hominis M. orale M. yeatsii
M. canis M. hyopneumoniae M. oxoniensis M. zalophidermidis
M. capricolum M. hyorhinis M. penetrans
M. citelli M. hyosynoviae M. phocidae
M. columborale M. imitans M. phocicerebrale
M. conjunctivae M. iowae M. pirum
M. cricetuli M. lacerti M. pneumoniae
Ureaplasma
U. canigenitalium U. diversum U. gallorale U. parvum
U. urealyticum


Mesoplasma
M. chauliocola M. florum M. photuris M. tabanidae
M. entomophilum M. grammopterae M. syrphidae  
Spiroplasma
S. cantharicola S. lineolae S. taiwanense S. citri
S. platyhelix      
Acholeplasma
A. laidlawii      

 

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200

Protocols

Manual (links to manufacturer's page) here

SDS (links to manufacturer's page) here

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200

References

Manual here (links to manufacturer's website)

SDS here (links to manufacturer's website)

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200

Applications & Benefits

Application Real-time PCR kit for detection of mycoplasma contamination in cell culture

Content Lyophilized Master Mix with FAM/HEX fluorescence probes, Internal Control and PCR-grade water Assays 25 or 50 (2 x 25) assays per kit

Shipping Ambient temperature Storage ≤ –15°C, store away from light

Shelf life 1 year (from date of delivery)

Product Group Mycoplasmafree Cell Culture

Sensitivity > 100 copies of the mycoplasma genome under optimal conditions (highly pure M. fermentans DNA, without Internal Control)

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200

Related products

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200