MycoStrip® 2.0 Mycoplasma Detection Kit: Rapid Mycoplasma Testing for Cell Culture

Mycoplasma are the smallest self-replicating organisms, at around 100 nm. They have no cell wall, so common antibiotics such as penicillin and streptomycin don’t touch them. They pass straight through standard 0.2 µm filtration and can’t be seen by eye or under a light microscope. That means a culture can look perfectly healthy while its biology is being rewritten.

Once established, mycoplasma compete with your cells for nutrients, alter metabolism and growth, change the expression of hundreds of genes and interfere with signalling and cytokine production. Their lipoproteins activate TLR2, which makes contamination especially damaging in immunology work with macrophages and other immune cells.

Mycoplasma by the numbers:

• Up to 35% of cell cultures are contaminated worldwide, including cell bank stocks
• 95% of contamination comes from just six species: M. orale, M. hyorhinis, M. arginini, M. fermentans, M. hominis and A. laidlawii
• 80% of lab operators carry mycoplasma, and talking or sneezing near cultures can spread it
• 50% of US labs don’t routinely test their cultures

Top reasons to test routinely:

• Protect precious cell lines, primary cells and clones
• Safeguard data reliability and reproducibility
• Meet journal requirements: most journals now ask for proof of mycoplasma-free cultures
• Avoid lost time, reagents and funding

MycoStrip® 2.0 detects contamination at around 1 CFU/ml, long before it reaches the levels where results are typically affected (approximately 10⁷ CFU/ml). InvivoGen recommends testing cultures every 2 to 3 weeks and quarantining all new cell lines until they test negative.

MycoStrip® 2.0 uses loop-mediated isothermal amplification (LAMP), a form of isothermal PCR run at a constant 65°C. A proprietary Reaction Mix, featuring a next-generation enzyme, amplifies a conserved region of the 16S rRNA gene found across Mollicutes, the class that includes Mycoplasma, Acholeplasma, Ureaplasma and Spiroplasma. The amplified DNA is then read on an immunochromatographic (lateral flow) strip. No DNA extraction, gel electrophoresis or thermal cycling program is required.

4 easy steps to detect mycoplasma in your cell cultures

1. Prepare your sample (~5 min): collect 1 ml of cell culture supernatant or FBS. Heating at 95°C for 5 minutes is optional but recommended to inactivate mycoplasma.
2. Incubate at 65°C for 30 minutes: combine the Reaction Mix, Reaction Buffer and sample. Extend to 50 minutes for maximum sensitivity (do not exceed 60 minutes).
3. Stop the reaction and load the cassette: add Migration Buffer and transfer the sample to the strip.
4. Read your results within 2 to 5 minutes: one band (control) means negative, two bands mean positive.

Just a heating device that holds 65°C (±5°C): a thermocycler, heat block, dry bath or water bath. MycoStrip® 2.0 has improved tolerance to temperature variation, so it performs reliably across standard lab equipment. Note that water baths can increase contamination risk.

Who is it for?

• Cell culture labs running routine contamination checks
• Core facilities and cell banks managing many lines
• Researchers preparing data for publication
• QC and quality assurance workflows in biotech and pharma research

MycoStrip® 2.0 is faster, more sensitive and more robust than the original MycoStrip®, thanks to an optimised high-performance enzyme.

Why GC/µl? Sensitivity is now reported in genome copies per µl, in line with European Pharmacopoeia edition 12.2, which favours molecular quantification. Based on in-house digital PCR studies, InvivoGen considers 1 GC/µl roughly equivalent to 1 CFU/ml for the most common contaminants.

Don’t stress. Precious cultures can usually be rescued with InvivoGen’s anti-mycoplasma reagents, also available from Millennium Science.

• Plasmocin® treatment: first-line elimination reagent that clears mycoplasma in about 2 weeks, with little to no toxicity to mammalian cells
• Plasmocure™: second-line option for Plasmocin®-resistant strains, using antibiotics with different mechanisms of action

After treatment (~2 weeks), re-test with MycoStrip® 2.0 and compare against your original sample. Keep an untreated backup vial so you can confirm the result or start again if needed. To prevent future contamination, consider adding Normocin™ or Plasmocin® prophylactic to routine culture media.

Get fast, clear mycoplasma testing for your cell cultures with MycoStrip® 2.0. Our team can help with pricing, pack sizes and the right workflow for your lab.

For research use only. Not for human or veterinary use. MycoStrip® is a trademark of InvivoGen.