简介：1PURPOSOFTHESTUDYThearticle99intheSpecificRulesforEnforcingtheFrontierHealthandQuarantineLawofthePeople’sRepublicofChinahasindicatedthatthehealthandquarantineorganizationshallpreventanyforeignersufferingfromvenerealdiseasesfromenteringtheterritory.Becausetheroutineconfirmatorytestforsyphilisfluorescenttreponemalantibodyab-

简介：Objective:Toevaluatetheclinicalutilityoftestingserumanti-treponemapallidumIgMantibodyinthediagnosisofsyphilispatients.Methods:Seventy-twocasesofsyphilisweretestedforspecificIgMantibodywithELISA,andtheresultswerecomparedwithRPRandTPPA.Results:ThesensitivityofIgMantibodywas73.3%(11/15)inprimarysyphilis,88.9%(16/18)insec-ondarysyphilis,andtherewasnosignificantdiffer-encebetweenthesevalues(x^2=1.6363,P>0.10).ThesensitivityofIgMantibodyindiagnosinglatentsyphi-liswasonly26.1%(6/23),muchlowerthanthedetec-tionrateinsymptomaticearlvsvDhilis(x^2=17.6189.P<0.005).RPRandTPPAwereboth100%sensitiveinlatentandearlysymptomaticsyphilis.Twowereposi,fiveforIgMinthe16caseswhohadreceivedregulartreatments2to24monthsbeforeenrolled.Conclusions:SpecificIgMantibodydetectiondoeesnotappearsuperiortoRPRandTPPAindiagnosingprimarysyphilis.ThediagnosisoflatentsyphilisshouldmainlyrelyonRPRandTPPA,sincetherearelowtitersofIgMantibodyatthatstage.IgMantibodytestingaloneshouldnotberecommendedformonitor-ingsyphilisdevelopmentortreatmentefficacy.Fur-therstudiesshouldbeconcerned.

简介：Objective:ToobtainrecombinantTreponemapallidumsubsp,pallidum(TP17KD)lipoproteininlargequantitiesbyamplificationandtofurtherpurifyantigensforlaboratorydiagnosisofsyphilisanddevelopmentofasyphilisvaccine.Method:TheTppl7lipoproteingenewasamplifiedfromtheTP(strainNichols),andthenitwasrecombinatedintoaplasmidpMAL-2candclonedwithinE.coli12-TB1.ThehostbacteriacontainingrecombinantplasmidswereinducedwithIPTG.TheTpp17KDlipoproteingenewasamplifiedbyus-ingPCRandpositivecloneswerescreenedwithdoubledigestionandPCR.RecombinantplasmidsweretransformedintoE.coliandtheE.colicarryingrecombinantplasmidswereinduced.TheexpressionofTP17KDwasdetectedbysodiumdedecylsulfate-polyacrylamidegelelectrophoresis(SDS-PAGE)andimmunoblot.Results:GelstainingwithCoomassieblueG-250showedthattheinducedE.colicarryingrecombinantplasmidcouldproduce60KDfusionproteinathighlevels.Gelscanningshowedthat17KDproteinexpressioninE.coliaccountedfor10%oftotalcellularprotein.Therecombinantproteinantigenreactedwiththeseraofsyphilispatients.Conclusion:Ourstudylaysacornerstonefordevelopingnewtechniquesoflaboratorydiagnosisforsyphilisandnewvaccines.Preliminaryclinicalapplicationshowedthatthefusionproteincouldbeusedforthediagnosisofsyphilis.

简介：Objectives:Todevelopamulti-nestedpolymerasechainreactioninanassaytodetectearlyTreponemapallidumandHaemophilusducreyiDNAintheswabsofgenitalulcers.Methods:Fourpairsofouterandinnerprimers,specifictothebasicmembraneproteingeneofTreponemapallidumandtothe16srRNAgeneofHducreyiweresynthesized.Themulti-nestedPCRwasdevelopedandappliedtodetectTreponemapallidumandHaemophilusdicreyiinclinicalswabs.Result:ThetwosamplesofstandardstrainsofHaemophilusducreyiandoneTreponemapallidumwereamplifiedandshowed309-bprRNAgeneofHaemophilusducreyiand506-bpDNAofTreponemapalidum,respectively.Outof51samplesofgenitalulcerdetected,29showedTreponemapallidumpositiveproductandnoHaemophilusducreyiDNAwasfound.Conclusion:Themulti-nestedPCRforTreponemapallidumandHaemophilusducreyicouldbeusefulforearlydetectionanddistinguishingdiagnosisbetweensyphilisandchancroid.

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简介：Objective:ToconstructtherecombinantplasmidcontainingGlycerophosphodiesterphosphodiesterase(Gpd)genefromTreponemapallidumandtransfectitintoHelacellstoexpresstheencodedoutermembraneprotein.Methods:TheGpdgenewasamplifiedfromthegenomicDNAofT.pallidumbypolymerasechainreaction(PCR)andinsertedintocloningvectorpUCm-T.TheinsertedGpdgenewassubclonedintotheappropriatesiteofpcDNA3.1(+)vector.Afteridentificationbysequencingandrestrictiveenzymesdigestion,therecombinantplasmidwastransfectedintoHelacellsusingliposomes.TheexpressedproteinwasidentifiedbyimmunocytochemistryandWesternblot.Results:ThetargetGpdgenesegmentwasapproximately1059bp.TheDNAsequenceoftheGpdgenecontainedinthepcDNA3.1(+)vectorwasconsistentwiththepublishednucleotidesequence.ThehomologyofthenucleotideandputativeaminoacidsequencesoftheGpdgenebetweenT.pallidumsubsp.pallidumNicholsandvariouspathogenictreponemalstrainsrangedfrom98%to100%.ImmunocytochemistryandWesternblotanalysisshowedthattheconstructedGpd-pcDNA3.1(+)vectorexpressedafusionproteinwithacalculatedmolecularmassof41KDainHelacellsandthattheexpressedproteinreactedwiththeserafromsyphilispatients.Conclusion:ThesuccessfulconstructionandexpressionoftheeukaryoticexpressionplasmidoftheGpdgenefromT.pallidumprovideapromisingtooltofurtherstudythebiologicalactivityofT.pallidumanddevelopaDNAvaccineforsyphilis.

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简介：Objective:ToevaluatetheclinicalapplicationofmultiplexPCRinthedetectionofTreponemapallidum,Herpessimplexvirus(HSV),andHaemophilusducreyi.Method:ThreestandardstrainswereusedtosetupamultiplexPCR(MPCR)fordetectingsyphilis,herpesgenitalis,andchancroidsimultaneously.Samplesfrom122patientswithgenitalulcerdisease(GUD)weresubjectedtoMPCRandtheresultswerecomparedwiththeseofdark-fiddmicroscopyandTPserology,HSVanligenELISA,andH.ducreyiculture,Result:Inthe122patientswithGUD,MPCRidentified34casesofT.palliduminfection,40casesofHSVinfection,and2casesofmixedinfectionofT.pallidumandherpes.NopositiveresultsofH.ducreyiwerefound.ThesensitivityofMPCRtoT.pallidumandherpeswas100%and93.3%,respectivdy.Thesensitivitiesofdark-fieldmicroscopyandTPserology,HSVantigenELISA,andH.ducreyiculturewas35.3%,50%and100%,respectively.Conclusion:MPCRshowedarelativelyhighersensitivityforT.pallidumascomparedwiththeroutinetechniques.AlthoughitssensitivityforHSVwasnotasgoodasthatofantigenELISA,italsoyieldedahighdetectionrate.MPCRcandetectmorethanonepathogen.Itissimple,quick,sensitive,andsuitableforclinicaluseorepidemiologicalinvestigation.

简介：TocloneandexpresstherecombinantoutermembraneproteinTp0453ofTreponemapallidumandtoanalyzetheimmuno-reactivityandimmunogenicityoftheexpressedprotein,theimmuno-dominantepitopeoftheTp0453wasamplifiedfromthecompletegenomeofT.pallidumbyPCR,subclonedintoexpressionvectorpQE32togeneratetherecombinantplasmidpQE32/Tp0453,thenexpressedinE.coliM15andanalyzedbySDS/PAGEandWesternblotting.ThefusionproteinexpressedwaspurifiedwithNi-NTAaffinitychromatography.Itsimmuno-reactivitywasassayedbyindirectELISA,andtheimmunogenicitywasdeterminedbyimmunizationwiththisfusionproteininNewZealandrabbits.Inthepresentstudy,afusionproteinofmolecularweightabout32kDawasobtained.AsdemonstratedbyWesternblotting,therecombinantproteincouldreactspecificallywithpositiveIgGseraofpatientswithsyphilis,andtheantibodiesagainstT.palliduminhumanseraweresuccessfullydetectedbyindirectELISA.BoththesensitivityandspecificityofELISAbasedontheTp0453fusionproteinaswere100%(30/30)whendetectedwithcontrolsera.IncomparisonwiththeresultsofIgGELISAwiththoseofTPPA.ItwasfoundthatthesensitivityofELISAwas96.8%andthespecificitywas100%.ThedifferenceofELISAandTPPAwasnotsignificant,andtheconcordanceofresultsbetweenELISAandTPPAwas98.2%.Inaddition,specifichumoralresponsescouldbeelicitedbyimmunizationwiththerecombinantfusionproteininNewZealandrabbitswithaspecificantibodytiterof1:1280after3successivedosesofimmunization.Theseresultsdemonstratethattheexpressedrecombinantfusionproteinshowsexcellentimmuno-competenceandprovidefoundationtodevelopaquickdiagnostickidappliedtodetectthepresenceofT.palliduminfections.

简介：Objective:Todevelopasensitive,specificandsimplemethodfordetectionofextremelylownumbersofT.palliduminclinicalspecimens,asasignificantadditiontotheserologictestsforsyphilisdiagnosis.Methods:Double-tubenestedPCR(DN-PCR)andsingle-tubenestedPCR(SN-PCR)assayswereperformedtoamplifyspecificfragmentsoftheDNApolymeraseIgene(polA)ofT.pallidum.SensitivityandspecificityofthetwoPCRassaysweretested.EightysixwholebloodspecimensfrompersonswithsuspectedsyphilisweredetectedbythetwonestedPCRmethods.TheTPPAtestwasusedasacomparisonfordetectingsyphilisinserafromcorrespondingpatients.Results:OnlyspecificampliconscouldbeobtainedduringamplificationoftheT.pallidumpolAgeneandthedetectionlimitwasapproximately1organismwhenanalyzedongelbythetwoPCRmethods.Of86clinicalspecimens,62werepositivebyTPPA.Ofthese,54and51werepositivebytheDN-PCRandSN-PCR,respectively,whichdoesnotrepresentastatisticallysignificantdifferencebetweenthetwoPCRtests.Of24TPPA-negativespecimens,5werepositivebybothDN-PCRassayandSN-PCRassay.Conclusion:TheSN-polAPCRmethodisextremelysensitive,specificandeasytoperformfordetectinglownumbersofT.palliduminclinicalbloodspecimensasacomplementarytoserologyforsyphilisdiagnosis.

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