Early monitoring of the human polyomavirus BK replication and sequencing analysis in a cohort of adult kidney transplant patients treated with basiliximab.docVIP

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Early monitoring of the human polyomavirus BK replication and sequencing analysis in a cohort of adult kidney transplant patients treated with basiliximab.doc

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Early monitoring of the human polyomavirus BK replication and sequencing analysis in a cohort of adult kidney transplant patients treated with basiliximab

Anzivinoetal.VirologyJournal2011,8:407 /content/8/1/407 RESEARCH OpenAccess EarlymonitoringofthehumanpolyomavirusBK replicationandsequencinganalysisinacohortof adultkidneytransplantpatientstreatedwith basiliximab ElenaAnzivino ,AnnaBellizzi,AnnaPaolaMitterhofer,FrancescaTinti,MarioBarile,MariaTeresaColosimo1, 1 1 2 2 2 4 1,5 1 3 3 DanielaFioriti ,MonicaMischitelli ,FernandaChiarini,GiancarloFerretti ,GloriaTaliani and ValeriaPietropaolo1,6* Abstract Background:Nowadays,betterimmunosuppressorshavedecreasedtheratesofacuterejectioninkidney transplantation,buthavealsoledtotheemergenceofBKV-associatednephropathy(BKVAN).Therefore,we prospectivelyinvestigatedBKVloadinplasmaandurinesamplesinacohortofkidneytransplants,receiving basiliximabcombinedwithamycophenolatemofetil-basedtripleimmunotherapy,toevaluatethedifference betweenBKVreplicationduringthefirst3monthspost-transplantation,characterizedbythenon-depletingaction ofbasiliximab,versusthesecond3months,inwhichthemaintenancetherapyactsalone.Wealsoperformed sequencinganalysistoassesswhetheraparticularBKVsubtype/subgrouportranscriptionalcontrolregion(TCR) variantswerepresent. Methods:WemonitoredBKviruriaandviremiabyquantitativepolymerasechainreaction(Q-PCR)at12hours(Tx), 1(T1),3(T2)and6(T3)monthspost-transplantationamong60kidneytransplantpatients.Sequencinganalysiswas performedbynested-PCRwithspecificprimersforTCRandVP1regions.Datawerestatisticallyanalyzedusingc2 testandStudent’ st-test. Results:BKVwasdetectedatTxin4/60urineandin16/60plasma,withmedianviralloadsof3,70logGEq/mL and3,79logGEq/mL,respectively,followedbyasignificantincreaseofbothBKV-positivetransplants(32/60)and medianvaluesofviruria(5,78logGEq/mL)andviremia(4,52logGEq/mL)atT2.Conversely,asignificantlydecrease ofpatientswithviruriaandviremia(17/60)wasobservedatT3,togetherwithareductionofthemedianurinary andplasmaviralloads(4,09logGEq/mLand4,00logGEq/mL,respectively).BKVTCRsequenceanalysisalways showedthepresenceofarchetypalsequences,withafewsingle-nucleotide