Testing for Mycoplasma species usingPolymerase Chain Reaction Techniques
All regulatory guidelines speciy that products manuactured using cells must be testedor the presence o Mycoplasma species. This includes all biologics and some vaccines.The testing is prescribed or master and working cell banks, virus seed lot and bulkharvests. Traditionally the tests or Mycoplasma have included cell culture or growth onmedia (broth and agar) however nucleic acid amplifcation techniques (NAT) may beused as an alternative to one or both o the other methods ater suitable validation bycertain guidelines. The reason or the acceptance o the NAT techniques by the EuropeanPharmacopoeia (EP) is that they have recognised that some samples are difcult to test orMycoplasma either due to cytotoxicity or due to the rapid turnaround o samples requiredor particular products.
Polymerase Chain Reaction
2.6.21 o the European Pharmacopoeia statesthat Nucleic acid amplifcation, such as PCR, techniques
may be used or detection o mycoplasmas. NAT indicatethe presence o a particular nucleic acid sequence andnot necessarily the presence o viable mycoplasma. Thispresents some advantages and disadvantages; theprincipal advantage is the sensitivity and specifcity othese assays. The main disadvantage is the detection onon viable Mycoplasma sequences and these sequencesmay be present in many o the reagents and culturematerial used in the production process. This is because,whilst the manuacturing o the reagents will haveeliminated the viable organisms, some DNA will remaindue to it being very difcult to remove. A number odierent techniques are available which can be appliedto detect this presence and the EP does not speciy aparticular method but does outline the validation whichis expected. The EP also indicates that commercial PCRdetection kits will be suitable or use or release assaysand specifes that certain elements o the validation maybe carried out by the manuacturer to acilitate use othe assay commercially. The PCR technique may be usedinstead o the culture method and the indicator cell culturemethod ater suitable validation as a release test, thussaving time and cost.
BioOutsource Service Offering
BioOutsource is pleased to oer the Applied BiosystemsPrepSEQ
detection kit or use in detectingMycoplasma to EP requirements. The PrepSEQ kit was