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  • 简介:摘要:医疗技术在经过不断发展后,带动了DNA检测技术的发展,并广泛的对其进行了应用,成为了法医鉴定检验方式中最主要的一种。法医所检验的DNA其实就是检验DNA在人体内的序列及其长度,进而对个人进行识别或是进行亲子鉴定。该方式是最为重要的法医物证鉴定方式之一。因此本文概述了DNA技术在法医鉴定中的应用,分析了DNA检测技术在法医鉴定中存在的问题,并对能够提高质量和控制水平的措施进行了提出,以此来促进法医鉴定事业在我国的发展。

  • 标签: 法医DNA检验 问题 尽量控制
  • 简介:Inordertoanalyzethesequencesoftheinternaltranscribedspacer(ITS)includingthe5.8SribosomalDNA(rDNA)ofcommondermatophytes,soastoobtainarapidandaccuratemethodtoidentifythespeciesofdermatophytesandtoestablishthephylogenetictreeofthesespeciestounderstandtheirrelationship,16strainsofdermatophyteswerecollectedandpreliminarilyidentifiedbymorphologicalcharacteristics.GeneralprimersforfungiITS1andITS4wereusedtoamplifytheITSrDNAofeachstrainswithPCR.ThePCRproductsafterpurificationweresequenceddirectlyandwereanalyzedthroughinternet.Intheresults,11strainswereidentifiedbymeansofmorphologicalfeatures,amongwhich5strainswereTrichophyton,5strainswereMicrosporumand1wasEpidermaphyton,whichwasconsistentwiththeresultsbymolecularbiology.Inthe5unidentifiablestrains,1strainwasprovedtobeChrysosporiumbymolecularbiology.Thesestrainsstudiedcouldbedividedinto3differentclassesasindicatedintheanalysisofthephylogenetictreeofthesequencesinITS,whichwerequitedifferentfromthoseofmorphologicalclassification.ItisevidentfromtheaboveobservationsthatthemolecularmethodofanalysisontheITSsequencesisarapid,highlysensitiveandaccurateapproachforthedetectionofdematophytespecies,however,itstillexhibitssomelimitationsneedingthesupplementationwithmorphologicalidentification.

  • 标签: 同源性 核醣体 脚癣 ITS序列 脚气
  • 简介:【摘要】随着医疗科技的不断发展, DNA甲基化检测方法逐渐增多, DNA甲基化属于遗传外饰物,是遗传学中的重要内容,对于该物质的研究一直是临床中的热点,但采用操作简单,能够有效帮助各界医学专家了解杂种优势以及基因学的秘密。为了临床研究提供准确依据,本文现对 DNA甲基化检测方法的临床研究进展展开综述,详情内容如下:

  • 标签: DNA 甲基化 检测方法 综述
  • 简介:TostudytheexpressioncharacteristicofJapaneseencephalitisvirus(JEV)prMEandEproteinsandtheefficacyofDNAimmunizationbydifferentrecombinantplasmidscontainingJEVprME(2001bp)andE(1500bp)genes,tworecombinants(pJMEandpJE)containingJEVprMEandEgenesfusedwithFLAGwereconstructedandthentransfectedintoHepG2andCOS-1cellsbylipnsomefusion.TheexpressionfeatureofFLAG-prME(about72kDa)andFLAG-E(about54kDa)proteinsintransfectedcellswereanalyzedbyWesternblotandtwoantibodysystems(anti-FLAGandanti-E).BALB/cmicewereimmunizedwith100μgoftwokindsofrecombinantsbyintramuscularinjection,andJEVJaGAr-01strains(10^5PFU/100μl)weregiventoBALB/cmicebyintraperionealinjection3wkaftertwiceDNAimmunizationbyalethalviruschallenge.BALB/cmicewereobservedfor21daysafterchallenge.80%plaquereductionneutralizationtestwasperformedtotitrateneutralizationantibodybeforeandafterviralchallenge.ItwasfoundthattheexpressionofproteinsassociatedwithpJMEandpJEwasdeterminedintransfectedcellswithanti-FLAGandanewproteinof11kDawasdetectedinHepG2andCOS-1cellstransfectedwithpJME.OnlyE(53kDa)proteinwasidentifiedastransfectedwithpJMEusingantiE.HigherlevelofneutralizationantibodiesandtheefficacyofprotectiveimmunitywereinducedwithpJMEimmunization,andweresimilartothoseinducedbyinactivatedJapaneseencephalitisvaccine,butwerebetterthanthoseinducedwithpJE.ItconcludesthattheexpressionlevelfromprMtoEproteinsofJEVisdifferentinvitro,andtheinvitroexpressionefficiencyofpJMEwasbetterthanthatofpiE.FLAG-prMEproteinexpressedbypJMEcouldbecleavedbypeptidasefromhost.TheefficacyofDNAimmunizationiscorrelatedtotheexpressioncharacterizationofrelatedproteinsexpressedinvitro.

  • 标签: 日本脑炎病毒 prME E蛋白 基因编码 基因重组构建 DNA免疫试验
  • 简介:AThepurposeofthisinvestigationwastostudythetherapeuticeffectofLamivudineonHBVDNAinperipheralbloodmononuclearcells(PBMC)andserum,andthelevelofcytokinesinserumofthepatientswithchronichepatitisB.Thepatientsweredividedintotwogroups(A=47,B=34),andtreatedbyLamivudine,routinemedicine,respectively.ThelevelsofHBV-DNAinPBMCandserumandcytokineswerealldetectedbeforeandaftertreatment.AfterthetreatmentofLamivndinefor36weeks,thetotalconversionnegativeratesofHBV-DNAinPBMCandserumofthepatientstreatedwithLamivudinewere55.32%(26/47)and61.70%(29/47),respectively.ThetotalnegativeconversionratesofHBV-DNAinPBMCandserumofthepatientstreatedbyroutinemedicinewere26.47%(9/34)and32.35%(11/34),respectively.TherewassignificantdifferencebetweenLamivudinegroupandroutinemedicinegroup(P<0.01).ThenegativeconversionratesofHBeAginserumofthepatientswere46.81%(22/47)and68.09%(32/47)attheendof24weeksand36weeks,andwerehigherthanthoseofroutinemedicinegroup(P<0.05andP<0.01).Thelevelsofalanineaminotransferase(ALT),aspartateaminotransferase(AST),ALT/ASTinserumofthepatientsafterbeingtreatedbyLamivudine,routinemedicineweredown-regulatedto(30.1±9.6)U/ml,(32.3±10.7)U/ml,0.9±0.1and(48.4±10.7)U/ml,(44.7±11.0)U/ml,1.1±0.2.Aftertheanalysisofvariance,thehighsignificantdifferencewasobviousbetweenthetwogroups(P<0.01).ItwasduetothehighlevelsofIL-6,IL-8andTNF-αinchronichepatitisBwhichcouldbedown-regulatedto(250.5±33.3)pg/ml,(153.4±22.2)pg/ml,(232.6±21.2)pg/mlbyLamivudine,whichwasmoreobviousthanthatofroutinemedicine(P<0.01).LamivudinehashightherapeuticeffectonthetreatmentofHBVDNAinPBMCandserumandhasbettertherapeuticeffectthanthatofroutinetherapy.Lamivudinemayalsohavehigherdown-regulatedinflammatoryinfiltrationandsecretioninlocalsitecaused

  • 标签: 细胞因子 乙型肝炎病毒 诱导因素 临床
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  • 简介:TheaimofthisstudyistoinvestigatethefeasibilityandmechanismofhIL-2-preSDNAvaccineaspreventionandtherapeuticapproachagainstHepatitisB.EukaryonexpressionvectorinvolvinghIL-2andpreSgenewasconstructedwithrecombinanttechniqueandtransferredintonormalBALB/cmiceandHBVtransgenicmice(Tg-Mice)respectively.Tnenaseriesofdetectionwereperformed:detectionofanti-preS2,HBsantibodyandHBsAginBALB/cmiceandTg-micewithELISA,quantificationofHBVDNAcopiesinHBVTg-miceserumwithreal-timePCR,determinationofhepatitisdegreewithimmunopathologicalHEstaininganddetectionofliverfunction.Anti-preS1canbedetectedat4^th,6^thand10^thweekininoculatedBALB/cmice.Injectionwithgenegungainedanadvantageovermuscularandsubcutaneousinjectionsinceitacquiredjust1/10inoculationquantity(10μg/mouse).HighestexpressionofIgG2aat4^thweeksuggestedThl-mediatedimmuneresponse,whichfacilitatedHBVcleaning.OfallinoculatedHBVTg-mice,80%ofthemshowedanfi-preS2,HBsantibodypositiveandHBVDNAdecreased,and20%showednegativeforHBsAg.HEstainingtohepatictissueshowedobviousinfiltrationofinflammatorycells,swellingandgranulardegenerationofhepatocytes.Inourstudy,IL-2-preSDNAvaccinewhichcanprovokethehumoralandcellularimmuneresponseandbreaktheimmunetolerancesupportsthedesignationandconstructionofnewvaccineagainstHBVandspecificimmuneremedyforHBVcontinuousinfection.

  • 标签: 疫苗接种 乙型肝炎病毒 病毒感染 hIL-2-preS DNA 小鼠
  • 简介:【摘要】目的:讨论乙肝病毒性肝炎患者临床医学检验应用探究。方法:选取我院治疗的乙肝病毒性肝炎的患者50例,均实行两对半和HBV-DNA定量检测,回顾性分析检测结果,选取的患者均需要使用两对半临床医学检验,采集患者的清晨空腹状态下静脉血4毫升,将血液放入离心机中,使用每分钟3000转,10分钟后,将血清分离出来,放在4摄氏度的条件下保存。结果:所有患者经临床检测诊断结果均为乙肝病毒性肝炎,其中小三阳患者占18例(36.0%),大三阳患者占13例(26.0%),其它类型占19例(38.0%),三组之间的差异性较为显著(P

  • 标签: 乙肝病毒性肝炎 两对半临床医学检验 HBV-DNA定量检测,应用探究
  • 简介:ThepurposeofthisstudywastoconstructaneukaryoticDNAvectorencodingamultipleepitopeantigen(MFC)ofhepatitisCvirus(HCV)andahepatitisBsurfaceantigen(HBsAg),andexploretheeffectofHBsAggeneontheimmunityofHCVmultiple-epitopeDNAconstructinvitroandinvivoinmice.AnHCVDNAvector(pVAX1-HBs-MFC)wasconstructedbyfusingHBsAggenetotheNterminalofanHCVmultiple-epitopeantigengene.ThepVAX1-HBs-MFCwastransfectedintoHEK293TcellsanditsexpressionwasmeasuredbyELISAandWesternblotting.BALB/cmicewereintramuscularlyimmunizedwiththepVAX1-HBs-MFC,andanELISAapproachwasappliedtodeterminethespecificantibodytitersandsubtypesinthemouseserum.Thecross-reactivityoftheantibodieswasalsocheckedwithtwosynthesizedHCVhypervariableregion1(HVR1)peptides.TheIFN-γproductionandcellproliferationofthemousespleencellswereevaluatedbyELISAandMTS(3-[4,5-dimethylthiazol-2-yl]-5-[3-carboxymethoxyphenyl]-2-[4-sulfophenyl]-2H-tetrazolium,innersalt)assays,respectively.TheexpressionofpVAX1-HBs-MFCwasdetectableinthetransfectedHEK293Tcells.TheserumantibodyresponsewaseffectivelyelicitedinBALB/cmiceinjectedwithpVAX1-HBs-MFC.ThehighesttiterofantibodyagainstHCV(MFC)was1:1280,andtheratioofIgG2a/IgG1was1.50±0.12atthefifthweekafterfirstimmunization.Moreover,thecollectedmouseserumantibodyhadtheabilitytocross-reactwiththetwosynthesizedHCVHVR1peptides.ThestimulationindexofthemousesplenocytestoMFCwas1.79±0.07,andtheIFN-γlevelwas287±6pg/mlatweek21afterfirstimmunization.ThehighesttiteroftheantibodyincontrolBALB/cmiceimmunizedwithpVAX1-MFCwas1:320,andtheratioofIgG2a/IgG1was1.33±0.11atweek5post-immunization.Furthermore,thestimulationindexofthemousesplenocytescellstoMFCwas1.52+0.06,andtheIFN-γlevelwas225±9.3pg/mlatweek21post-immunization.TheHBsAggenecanenhancetheeffectsofanHCVmultiple-epitope

  • 标签: 乙型肝炎病毒 丙型肝炎病毒 免疫反应 DNA 抗原决定基
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  • 简介:TherRNAgeneticlocusisfoundinallprokaryoticorganisms,andishighlyconservative,althoughitsrelativelystablevariationsarefoundfrequentlyindifferentbacteria.Theutilityofthislocusasataxonomicandphylogenetictoolhasbeenreportedwidely.Thisstudy,aimedat16SrRNAgene(16SrDNA)andwiththehelpofbiomolecularmethods,attemptedtoachievethegoalofrapididentificationofcommonpathogensInthisstudy,333clinicalisolatedpathogenicbacteriawerecollected。TwopairsofprimerswerechosenandlabeledwithdifferentfluorescentdyesandthenusedtoamplifythegenomicDNAextractedfrombacteria.ThePCRproductswerethendetectedbycapillaryelectrophoresis-singlestrandconformationpolymorphism(CE-SSCP).Inordertopursuehigherresolutionandpeak-separationeffect,ahighefficientseparatingmedium,linerpolyacrylamidedel(LPA),wasputtouseinthisstudy.Finally,everybacteriacolonygenerateddistinctpatternsfromeachother,whichwereeasilytobeusedforidentification.TheseresultsindicatedthatPCR-CE-SSCPwasarapididentificationmethodforbacterialidentification,withtheaspectsofhighefficiencyandhighprecision.Comparedwithtraditionalmethod,thistechnologyisofgreatutilityforclinicaluseespeciallyforitshighsensitivity.

  • 标签: 毛细电泳 细菌检测 检测方法 DNA 荧光 PCR
  • 简介:【摘要】目的:对肺结核合并乙型肝炎患者应用抗结核药物治疗的临床价值进行系统研究,以证实其对肝功能与HBV-DNA水平的影响。方法:选择我院2019年01月~2020年01月收治的50例肺结核合并乙型肝炎患者作为实验对象,根据血清HBV-DNA水平分组后探究每组治疗效果。对照组(25例):低水平组;实验组(25例):高水平组。对比两组患者肝功能损伤时间、肝功能恢复时间、肝损害率。结果:实验组肝功能损伤时间为(15.41±3.88)d、肝功能恢复时间为(84.14±6.35)d,较之于对照组,差异具备统计学研究意义(P

  • 标签: 抗结核药物 肺结核 乙型肝炎 肝功能 HBV-DNA水平