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1、JournalofAssistedReproductionandGenetics,Vol.15,No.5,1998ComparisonofFISH,PRINS,andConventionalandFluorescentPCRforSingle-CellSexing:SuitabilityforPreimplantationGeneticDiagnosisIANFINDLAY,1,4NICOLECORBY,2ANTHONYRUTHERFORD,3andPHILIPQUIRKE2Submitted:December15,1997preimplantationgeneticdiagno
2、sis(PGD),itwasshownAccepted:January8,1998tobeunreliableand,moreimportantly,inaccurate,duePurpose:Althoughconventionalpolymerasechainreactiontoamplificationfailure(1),However,PCRhasbeen,(PCR)wasthefirstmethodusedforsexinginpreimplanta-andstillis,usedsuccessfullytodiagnosecysticfibrosistiongene
3、ticdiagnosis,fluorescentinsituhybridizationstatus(2-4)aswellasothersingle-genedefectsin(FISH)hasbecomethemethodofchoice.RecentlytwonewPGD.techniques,primedinsitusynthesis(PRINS)andfluorescentFluorescentinsituhybridization(FISH)hasnowPCR,havebeendeveloped.Thisstudycomparesthereliabil-beenadopt
4、edbymostPGDcentersworldwideastheityandaccuracyofthesefourtechniquesinsinglecells.methodofchoiceforsexing(5-8).Results:Inbuccalcells,fluorescentPCRandFISHhadTheapplicationofFISHinPGDhasshownthatsimilarreliability(94and93%)andaccuracy(97and96%)chromosomalmosaicismiscommonatthecleavagerates.Ther
5、eliabilityandaccuracyofPRINS(91and25%)stageofdevelopment(6).ThisfindinghasveryandconventionalPCR(79and89%)werelower.Inhumanblastomeres,FISHandflourescentPCRhadsimilarreliabil-importantimplicationsfordiagnosisofdominantsin-ity(100%,717;95%,190/201)rates.Accuracyratesweregle-genedisorders,monos
6、omies,andtrisomiesaswell71%(5/7)and99%(188/190)forFISHandfluorescentPCR,asfortheunderstandingofearlyhumandevelopment.respectively,however,toofewblastomereswereanalyzedbyThesestudieshavenowbeenconfirmed(9,10),demon-FISHformeaningfulcomparison.However,whenthesedatastratingthatembryoaneuploidyan
7、d/ormosaicism,atarecomparedwithpublisheddata,themethodofchoiceforleastinhumans,occursatarateof40-80%,dependingblastomeresexingappearstobefluorescentPCR.onthenumberofFISHprobesandembryoquality.Conclusions:FlouroscentPCRhasmajorimplicationsThis
