10CFU™ Sensitivity Standards

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 in an 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 highest sensitivity by PCR-based methods. After extraction the 10CFU™ extract can be directly used for PCR.

Recommended Use

10CFU™ Sensitivity Standards are intended for validating the robustness and sensitivity of NAT-based mycoplasma tests.

Content

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. fermentas, 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. 102-1003 Mycoplasma arginini
• Cat. No. 102-2003 Mycoplasma orale
• Cat. No. 102-3003 Mycoplasma gallisepticum
• Cat. No. 102-4003 Mycoplasma pneumoniae
• Cat. No. 102-1103 Mycoplasma salivarium
• Cat. No. 102-5003 Mycoplasma synoviae
• Cat. No. 102-6003 Mycoplasma fermentans
• Cat. No. 102-7003 Mycoplasma hyorhinis
• Cat. No. 102-8003 Acholeplasma laidlawii
• Cat. No. 102-9003 Spiroplasma citri
• Cat. No. 102-0002 Mycoplasma Set

Result Evaluation

Application of a PCR-based mycoplasma test, e.g. Venor^®GeM qEP Mycoplasma Detection kit

Required Consumables

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.