Mycoplasma Testing - Detection & Elimination
Mycoplasma Testing
Mycoplasma Detection & Elimination Kits
Visually detect Mycoplasma by eye in 1 hour!
Mycoplasma Detection Kit
| Rapid | Detect Mycoplasma in 1 hour by visual colour change! | |
| Simple | Add-Incubate-Detect protocol with results easily read by eye meaning no expensive equipment required. | |
| Sensitive | Detect as little as 500cfu Mycoplasma per 1ul of cell culture supernatant. | |
| Flexible | Detect 28 species of mycoplasma in adherent & suspensions cells such as Vero, MDCK, SP2/0, 293T, HepG2. HeLa, A549, MB-MDA231, L929, MEF, CHO, NS0, 293F, mouse hybridomas, Sf9, BHK21 & more. | |
| Compatible | With a wide selection of cell culture media & sera such as Fetal bovine/calf serum, horse serum, Gibco KSR serum replacement & CD FortiCHO, CDM4, Expi 293 Medium, CD Hybridoma, Grace, DMEM, 1640, F12 & more. |
MycoPlasma Elimination Kit
| Effective | Remove mycoplasma through membrane disruption allowing for the elimination of even antibiotic-resistant mycoplasma | |
| Complete | Eliminate intracellular and extracellular mycoplasma in 3 days! | |
| Safe | Non-toxic to cells | |
| Validated | In commonly used cell lines including mouse and human embryonic stem cells, iPS cells, HEK293, Hela, HepG2, HCT116, COS-7, Vero, Huh-7, MDCK, PANC-1, SW620 and U2OS. |
| MycoGenie MycoPlasma Elimination Kit | |
|---|---|
| Size: | 100 or 500µL |
| Storage: | -20°C |
| Assay Time: | 3 - 7 days |
Validation Data
At Assay Genie we have published a validation white paper outlining how our Mycoplasma Detection Kit matches the Gold Standard of qPCR Mycoplasma detection!
Mycoplasma Detection & Elimination FAQs
At Assay Genie we have compiled a list of some commonly asked questions regarding the MycoGenie Mycoplasma Detection & Elimination Kits along with their solutions!
FAQs
1.) What is the sensitivity of the MycoGenie Rapid Mycoplasma Detection Kit?
The MycoGenie Rapid Mycoplasma Detection Kit is suitable for most cell culture experiments. It can accurately detect at least 500 cfu of mycoplasma from 1 μl of culture supernatant (5 × 105 cfu/ml). Typically, the mycoplasma content in culture supernatant is between 106 -108 cfu/ml. According to published literature, one single mycoplasma in cell culture can grow to 106 cfu/ml in 3-5 days. Therefore, it is highly recommended to detect after the third day after cell passage or after replacing media.
2.) What to do if the detection color changes immediately after supernatant is added or different colors (not blue or purple) appear during the reaction?
In rare cases, ingredients in the media interfere with the color of the MycoGenie reagent. For example, Cell Boost 5 (Hyclone) makes the MycoGenie reagent appear pink. To avoid this:
| Steps | Description |
| 1. | Collect a small amount of culture supernatant or cell suspension and centrifuge at 500 × g for 5 |
| 2. | Centrifuge again at high speed (> 12,000 × g) for 5 min to precipitate mycoplasma in the supernatant. Discard most of the supernatant and leave about 50μl in the tube. Add 950μl of sterile water and mix gently by pipetting. |
| 3. | Repeat Step (2) for three times. Discard most of the supernatant and leave about 50μl in the tube. |
| 4. | Take 1μl of supernatant for detection. |
3.) How to protect cells from mycoplasma contamination?
If mycoplasma contamination occurs, it is recommended to discard the cells to prevent other cells from contamination. If a mycoplasma positive result is detected, the same batch of cells should be discarded. Also, consider MycoGenie MycoPlasma Elimination Kit (Cat. No. MORV0195) as an excellent choice to remove mycoplasma from cell culture.
4.) How to avoid false positives?
Generally, no false positives will occur during proper usage. The reaction tube should never be reopened, due to false positives resulting from aerosol contamination. Change tips between samples and always add the positive control last.
5.) How many mycoplasma species can be detected by MycoGenie Rapid Mycoplasma Detection Kit?
There are 28 species of mycoplasma that can be detected accurately by MycoGenie Mycoplasma Detector:
A. laidlawii*
M. hominis*
M. arginini*
M. fermentans*
M. hyorhinis*
M. salivarium*
M. pirum*
M. orale*
A. granularum
M. alkalescens
A. pleciae
M. neophronis
M. timone
M. caviae
M. alvi
M. bovis
M. primatum
M. leopharyngis
M. maculosum
A. oculi
M. iners
M. gallinarum
M. sphenisci
M. bovigenitalium
M. auris
M. columbinum
M. lipophilum
M. falconis
* More than 95% mycoplasma contaminations in cell culture are caused by these 8 kinds of mycoplasma
Related Resources
What is Mycoplasma?
Mycoplasma is a type of bacteria that lacks a cell wall and flexible membrane. The size of mycoplasma bacteria is typically 0.15-0.3µm. Due to its small size, mycoplasma can escape through filtering systems in cell culture facilities. Furthermore, these organisms are resistant to most antibiotics commonly employed in cell cultures such as such as penicillin and streptomycin.
Mycoplasma testing and elimination procedures help to avoid the costly consequences of a cell culture contamination and ensure safe biopharmaceuticals and reliable scientific results. At Assay Genie we have developed Mycoplasma Detection and Mycoplasma Elimination kits.
Schematic of Mycoplasma
Mycoplasma Background
Mycoplasma testing is required to detect the presence of mycoplasma contamination in biopharmaceutical laboratories. Mycoplasma contamination is one of the main problems in cell culture. Mycoplasma are ubiquitous in the environment. In cell culture laboratories, the majority of mycoplasma infections in continuous cell lines are caused by as little as six contaminant species of bovine, swine, and human origin. These microbial agents easily propagate from an infected cell culture batch to another via aerosol transmission or inadequate aseptic techniques. If undetected, mycoplasma contamination can have significant effects on the quality and reliability of your cell culture preparations.
Mycoplasmal infection can often dwell in cell cultures for a long period of time without any visible cell damage. Therefore, it is vital to routinely use effective detection methods to test for the presence of mycoplasma contamination. Mycoplasma contamination represents a serious issue for academic and biopharmaceutical laboratories due to their effects on cell properties and on safety of biological products.
Regular mycoplasma testing and control procedures help avoiding the costly consequences of a cell culture contamination and ensuring safe biopharmaceuticals and reliable scientific results.
How is Mycoplasma Eliminated?
One option for mycoplasma elimination is treatment with antibiotics, which do not damage or alter cells. Antibiotics such as penicillin, which attacks bacterial cell walls, are ineffective in this instance, since mycoplasma lacks a true cell wall. Several antibiotics eliminate mycoplasma effectively, such as: Tylosin, Neomycin, Tetracycline and Gentamicin. However, the efficacy of these antibiotics is restricted to specific mycoplasma species and frequently only reduce the concentration of mycoplasmas rather than disinfect the cell culture. Hence, as soon as treatment is concluded, contamination will recur.
Our more effective option for Mycoplasma Elimination is the MycoGenie MycoPlasma Elimination Kit. This kit is a very robust method for removing Mycoplasma in cell culture, serum, and medium. Unlike conventional antibiotics, this kit kills mycoplasma by disrupting the membrane structure, and can remove antibiotic-resistant mycoplasmas, so as to achieve more effective elimination of mycoplasma rather than inhibiting proliferation. This kit is also effective on Gram negative and positive bacteria.