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.