Chlamydiatrachomatisoutermembraneprotein2(Ctomp2)isamajorimmunogeninchlamydialinfectionsandahighlygenus-conservedstructuralproteinofallChlamydiaspecies.Topurifytheproteinandtopreparemonoclonalantibodies(mAbs)againstit,therecombinantproteinwasinducedbyIPTG,whichwasconfirmedbySDS-PAGEandpurifiedbymeansofaNi2+-chargedresincolumn.ThedenaturedproteinwasrefoldedintheGSH-GSSHbuffergraduallyandidentifiedbyWesternblotting.ThentheBALB/cmicewereimmunizedwiththerecombinantproteintopreparethemAbagainstCtomp2.TheobtainedmAbswerecharacterized.GenitalspecimensweretestedwithindirectELISAmostlymadeofthemAbandcellculturein84patientswithgenitalsymptoms.Theresultsshowedthathigh-levelexpressionoftherecombinantproteinwasachieved,whichexistedasinclusionbodyandamountedto38%oftotalbacteriumprotein.AmAbagainstCtomp2wasobtained.ItbelongstoIgG2b.Thetiterswereashighas1:40000.TheWesternblottingshowedthatthemAbonlyreactedwiththerecombinantprotein.IthadnocrossingreactionsagainstE.coli,N.gonorhoea,M.hominis,U.urealyticumandM.penetrans.Ithadhighspecifity.Incomparisonwithgoldstandardtest-cellculture,thesensitivities,specificities,positivepredictivevaluesandnegativepredictivevaluesofindirectELISAwere95.24%,100%,100%and98.44%,respectively.Theabove-mentionedresearchworkcontributednotonlytothefurtherstudyofthestructureandfunctionofthisprotein,butalsototheestablishmentofthemethodforitsclinicalapplication,forithadnotbeenreportedbefore.
