Mycoplasma detection and removal is critical for biologics produced for clinical studies and licensed products for human use. Testing for mycoplasma is one of the most complicated carried out for all microbes as the family has many different and often fastidious requirements for growth. BioOutsource offer mycoplasma testing services under cGMP and to the current ICH, European Pharmacopoeia and United Stated Pharmacopoeia guidelines.
There are two components to mycoplasma testing described within the EP; Agar and broth and the in vitro indicator cell culture method. The sample matrix to be tested should be qualified in the assay prior to testing; qualification involves spiking the test material with test organisms at low concentrations and demonstrating detection. The following organisms are suitable as test organisms: A.laidlawii, M.gallisepticum, M. fermentans, M.hyorhinis, M.orale, M.pneumoniae.
Agar and Broth
For the agar and broth method, the sample is inoculated into different liquid and solid media types which are subsequently incubated in different environmental conditions (aerobic and anaerobic). During the incubation period, samples from the inoculated broth are also taken and inoculated onto the agar growth media; the entire test lasts for 28 days.
In vitro Indicator Cell Culture
The in vitro indicator cell culture method requires that the sample is inoculated onto a suitable cell substrate (Mouse 3T3 or Vero cells), cultured and stained with a fluorescent dye that binds to DNA. The cultures are observed by light microscopy. BioOutsource is able to report the results of the assay with a Certificate of Analysis within 5 days of the final observations.
Rapid Mycoplasma PCR
The European Pharmacopoeia has now accepted the replacement of the traditional mycoplasma assays with a PCR based method following appropriate validation. Since 2012, the US Food and Drug Administration (FDA) has accepted the use of PCR Mycoplasma Detection as a release method for a biopharmaceutical product. This now allows manufacturers to replace the existing method which lasts 28 days with a rapid technique. This rapid Mycoplasma PCR can be performed at BioOutsource and a result reported in 48 hours from receipt of sample.
Contact our biosafety scientists to discuss your mycoplasma testing requirements.