The European Pharmacopoeia Chapter 2.6.7 (EP 2.6.7) and the Japanese Pharmacopoeia Chapter G3 (JP G3) describe nucleic acid amplification technology (NAT)-based tests for mycoplasma such as PCR. Both require to show the detection of 10 Colony Forming Units per ml of sample volume (CFU/ml), if said tests are proposed as alternative methods to traditional culture assays. Such sensitivity must be demonstrated as part of the robustness testing for each specific sample matrix of interest. However, since most cell culture labs and production facilities do not have access to a microbiology lab, culturing or handling viable mycoplasma colonies to use as reference tools for sensitivity testing is inadmissible.
Our 10CFU™ Sensitivity Standards contain irreversibly inactivated mycoplasma (amount corresponding to 10 CFU) and allow safe and reliable validation. Once resuspended in the sample matrix of interest (as in EP 2.6.7 / JP G3), these samples must be positively tested with the applied method. Due to the mycoplasma inactivation, the 10CFU™ Sensitivity Standards are not suitable for mycoplasma culture or culture-based methods. Extensive proficiency testing indicated that mycoplasma DNA extraction is indispensable to achieve the highest sensitivity by PCR-based methods. After extraction the 10CFU™ extract can be directly used for PCR.
10CFU™ Sensitivity Standards are intended for validating the robustness and sensitivity of NAT-based mycoplasma tests.
Unit package: 3 vials containing 10 CFU of the particular mycoplasma as well as 2 vials negative control
Set package: 2 vials of each EP 2.6.7-listed mycoplasma containing 10 CFU (18 vials in total: M. arginini, M. orale, M. gallisepticum, M. pneumoniae, M. synoviae, M. fermentans, M. hyorhinis, A. laidlawii, S. citri) as well as 2 vials negative control. M. salivarium is not included in the 10CFU™ Mycoplasma Set.
The components are lyophilized.
Available Species and Ordering Information
Cat. No. 102-1003Mycoplasma arginini
Cat. No. 102-2003Mycoplasma orale
Cat. No. 102-3003Mycoplasma gallisepticum
Cat. No. 102-4003Mycoplasma pneumoniae
Cat. No. 102-1103Mycoplasma salivarium
Cat. No. 102-5003Mycoplasma synoviae
Cat. No. 102-6003Mycoplasma fermentans
Cat. No. 102-7003Mycoplasma hyorhinis
Cat. No. 102-8003Acholeplasma laidlawii
Cat. No. 102-9003Spiroplasma citri
Cat. No. 102-0002Mycoplasma Set
Application of a PCR-based mycoplasma test, e.g. Venor®GeM qEP Mycoplasma Detection kit
Sterile 1.5 ml reaction tubes, filter tips, appropriate DNA extraction kit
Required Lab Devices
Microcentrifuge, pipetting equipment
Shelf Life and Storage
The date of expiry is given on each package. The unopened kit should be stored at +2 to +8 °C.
Figure - 10CFU™ Sensitivity Standards are resuspended in 1 ml cell culture medium, of which 200 µl are used for DNA extraction with the Venor®GeM Sample Preparation Kit. The mycoplasma PCR test is then performed with the Microsart® ATMP Mycoplasma kit and a Corbett RotorGene®6000 cycler.