MycoplasmatestsaccordingtotheEuropeanPharmacopoeia(EP)Chapter2.6.7requireasensitivityof10colonyformingunits(CFU)permlsamplevolumeforNAT-basedmethodssuchasPCRinordertoreplacetrADItionalculturemethods.Thissensitivitymustbeaccomplishedaspartoftherobustnesstestingforanyparticularsamplematrix.
Usingvitalmycoplasmaisnotacceptableforthemajorityofcellculturelaboratoriesduetosafetyregulations.Our10CFU™sensitivitystandardscontainnon-vitalmaterialandallowsafeandreliablevalidation.
ThemycoplasmaweregrowninculturemediumasdescribedinEP2.6.7,subsequentlytitratedandplatedonHayflickandFreymediumforCFUdetermination.Themycoplasmawereharvestedintheearlylogarithmicgrowthphasetoensureahighratioofvitaltonon-vitalmycoplasmaandthusalowratioofGU*toCFU.AllstrainswereobtainedfromtheNationalCollectionofTypeCultures(NCTC),UK,andcultivatedinlowpassages.
ScopeofApplication
10CFU™SensitivityStandardsareintendedforvalidatingtherobustnessandsensitivityofNAT-basedmycoplasmatests.
KitComponents
Unitpackage:3vialscontaining10cfuoftheparticularmycoplasmaaswellas3vialsnegativecontrol
Setpackage:2vialsofeachEP2.6.7-listedmycoplasmacontaining10cfu(18vialsintotal)aswellas2vialsnegativecontrol
Thecomponentsarelyophilized.
SpeciesandOrderInformation
- Cat.No.102-1003Mycoplasmaarginini
- Cat.No.102-2003Mycoplasmaorale
- Cat.No.102-3003Mycoplasmagallisepticum
- Cat.No. 102-4003Mycoplasmapneumoniae
- Cat.No.102-5003Mycoplasmasynoviae
- Cat.No.102-6003Mycoplasmafermentans
- Cat.No. 102-7003Mycoplasmahyorhinis
- Cat.No.102-8003Acholeplasmalaidlawii
- Cat.No. 102-9003Spiroplasmacitri
- Cat.No. 102-0002MycoplasmaSet
Evaluation
ApplicationofaPCR-basedmycoplasmatest,e.g.Venor®GeMqEPMycoplasmaDetectionkit
RequiredConsumables
Sterile1.5mlreactiontubes,filtertips,appropriateDNAextractionkit
Requiredlabdevices
Microcentrifuge,microPipettes
ShelfLifeandStorage
Thedateofexpiryisgivenoneachpackage.Theunopenedkitshouldbestoredat+2to+8°C.
Figure–10CFUwerere-sUSPendedin1mlcellculturemedium,200µlwereusedforDNAextractionwiththeVenor®GeMSamplePreparationKit.ThemycoplasmaPCR-testwasperformedwiththeMicrosart®ATMPMycoplasmakitandaCorbettRotorGene®6000cycler.