Myco-Visible Mycoplasma qPCR Detection Kit

The Myco-Visible Mycoplasma qPCR Detection Kit enables fast and highly sensitive detection of Mycoplasma, Acholeplasma, and Spiroplasma, including the species most frequently associated with cell culture contamination: M. arginini, M. fermentans, M. hominis, M. hyorhinis, M. pirum, M. salivarium, M. orale, and A. laidlawii. This assay employs a nucleic acid amplification technique (NAT) and complies with the European Pharmacopoeia (EP) 2.6.7, Japanese Pharmacopoeia (JP) 18 G3, and the United States Pharmacopeia (USP) <63>. It has been validated across a broad selection of real-time PCR platforms and provides reproducible and reliable performance. For EP/JP/USP-compliant testing, suitable DNA extraction kits include the MagBeads FastDNA Kit (Cat. No. 116575192), SPINeasy DNA Kit for Forensics (Cat. No. 116597050), and SPINeasy DNA Kit for Tissue (Cat. No. 116532050).

Compared with traditional culture-based testing, which can take up to 28 days, this assay offers a rapid and precise alternative and does not require viable mycoplasma organisms. Consequently, it supports detection of both culturable and non-culturable species. The method demonstrates a sensitivity of ≤10 CFU/mL, supporting regulatory-compliant testing across multiple sample types.

Results Interpretation:

Performance:

Table 1. Analytical sensitivity of the Myco-Visible Mycoplasma qPCR Detection Kit.
Limit of detection (LOD) was evaluated following EP 2.6.7, JP 18 G3, and USP <63> guidelines using pharmacopeia-listed species spiked into cell culture medium (100, 10, and 5 CFU/mL; n=24). The kit consistently achieved high detection rates across all species, with an LOD₉₅ of 5 CFU/mL, meeting the required sensitivity of ≤10 CFU/mL.

Table 2. Specificity of the Myco-Visible Mycoplasma qPCR Detection Kit.
Specificity was evaluated in accordance with EP 2.6.7 using high concentrations (1 × 10⁵ copies/reaction) of non-target bacterial and mammalian DNA. No amplification was observed for off-target species.

Application Notes

The Myco-Visible Mycoplasma qPCR Detection Kit is designed to detect mycoplasma contamination in cell cultures by amplifying the conserved regions of the mycoplasma genome using qPCR.

Usage Statement

Research Use Only (RUO)