简介:Objective:Todetectandquantitategenitalherpessimplexvirus(HSV)DNAinspecimensfrom100patientsclinicallydiagnosedwithgenitalherpes.Methods:PolymeraseChainReaction(PCR)andenzyme-linkedimmunosorbentassay(ELISA)wereusedwithastandardcurveofDNAcopiesofHSVasquantitativecontrast.Results:Ninety-threecaseswereconfirmedHSVpositiveand7caseswerefoundtobenegative.Therewere58casesofHSV-2(62.4%)and35casesofHSV-1(37.6%)amongthe93positivecases.ThenumberofDNAplasmidsrangedfrom115to1.1×l0^5per250pLamongthe93positivesamples(mean=7.1×10^4/250μL).ThenumberofHSVDNAplasmidsrangedfrom136to1.1×l0^5copiesper250pL(mean=7.6×10^4)amongthosewithHSV-2,and115to9.4×10^4per250pL(mean=6.3×10^4)amongthosewithHSV-1.Meanwhile10μLofextractedanddissolvedDNArandomlytakenfrom8eachofHSV-2andHSV-1samplesweretested.ThenumberofHSV-2DNAplasmidsrangedfrom35copiesto2.7×10^4(Mean=l.8×10^4)andthenumberofHSV-1DNArangedfrom29to2.5×10^4(Mean=1.6×10^4).Inthe7negativecases,thequantityofHSVplasmidswaszero.Conclusion:ThesensitivityofELISAquantitation(93%)isequaltothatofSouthernblot.ThesensitivityofPCRfordiagnosisis91%,and88%forPCRtyping.
简介:Objective:ToexploretherelationshipbetweenquantitativeTreponemapallidumDNA(TP-DNA)PCRtestingandtheToludineRedUnheatedSerumTest(TRUST)inpatientswithsyphilisbeforeandaftertreatment,andevaluatetheclinicalvalueofquantitativeTP-DNAtestinginthediagnosisandtreatmentevaluationofsyphilis.Methods:29patientswithprimary(12cases)orsecondary(17cases)syphilis,whometthecriteriasetforthisstudywererecruitedassubjects.Allpatientsweretreatedwith2.4millionunitsbenzathinepenicillinIMweeklyfor3weeks.QuantitativetestsofTP-DNAinthepatients'plasmawereperformedusingFQ-PCRbeforeandafterthetreatment.SerologictestsincludingTRUSTandTPPAwerealsoperformed.Results:Beforethetreatment,9outof12primarysyphilispatients(75%)andallsecondarysyphilispatients(17/17)testedpositiveforTreponemapallidum(TP)byTP-DNAtesting.TheaveragequantitativetestvaluesofTP-DNAinprimaryandsecondarysyphilispatientswere(3.38±2.34)×10^4and(5.73±1.33)×10^6copies/ml,respectively.Afterthreemonthsoftreatment,1ofthe9primaryand5outof17secondarysyphilispatientswerepositiveuponTP-DNAtesting,respectively.TheaveragequantitiesofTP-DNAwere2.01×10^2copies/mlinprimaryand5.87×10^2copies/mlinsecondarysyphilispatientswithpositiveTRUSTandTP-DNAtests,and3.09×10^2copies/mlforthosewithnegativeTRUST,respectively.Afterninemonthsoftreatment,alltheprimaryandsecondarysyphilispatientswerenegativeuponTP-DNAtesting,whileallprimaryand14of17(82.35%)secondarysyphilispatientsshowednegativeTRUSTresults.Conclusion:ThattheresultsofTP-DNAtestsarenotconsistentwiththoseofTRUSTbeforeandaftertreatmentindicatesthatquantitativeTP-DNAtestingmayhavevaluableclinicalsignificanceintheearlydiagnosisandevaluationoftreatmentregimensforsyphilis.
简介:Objectives:Todevelopamulti-nestedpolymerasechainreactioninanassaytodetectearlyTreponemapallidumandHaemophilusducreyiDNAintheswabsofgenitalulcers.Methods:Fourpairsofouterandinnerprimers,specifictothebasicmembraneproteingeneofTreponemapallidumandtothe16srRNAgeneofHducreyiweresynthesized.Themulti-nestedPCRwasdevelopedandappliedtodetectTreponemapallidumandHaemophilusdicreyiinclinicalswabs.Result:ThetwosamplesofstandardstrainsofHaemophilusducreyiandoneTreponemapallidumwereamplifiedandshowed309-bprRNAgeneofHaemophilusducreyiand506-bpDNAofTreponemapalidum,respectively.Outof51samplesofgenitalulcerdetected,29showedTreponemapallidumpositiveproductandnoHaemophilusducreyiDNAwasfound.Conclusion:Themulti-nestedPCRforTreponemapallidumandHaemophilusducreyicouldbeusefulforearlydetectionanddistinguishingdiagnosisbetweensyphilisandchancroid.
简介:目的对人类免疫缺陷病毒(humanimmunodificiencyvirus,HIV)感染者中合并丙型肝炎病毒(hepatitisCvirus,HCV)感染的情况进行流行病学调查和统计学分析,评估合并感染的患病率和相关影响因素。方法对各医院在2006—2008年收治的门诊和住院HIV感染病例,用统一的流行病学调查表进行登记;并进行HCV抗体、肝功能和CD4细胞检测;多元回归法分析HIV合并HCV感染风险因素。结果HIV感染者978例,合并HCV的感染率为33.9%,静脉吸毒途径感染HIV占合并感染者的81.3%,30~45岁年龄组HIV合并感染率为42.6%,无业人员HIV合并感染率为61.9%,均明显高于其他组。结论年龄30~45岁和无正当职业的人群HIV合并HCV感染率较高,静脉吸毒是HIV合并HCV感染的主要传播途径。
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.
简介:目的研制人乳头瘤病毒16型(HPV16)L1/E7CTL重组杆状病毒载体。方法以包含HPV16标准病毒株的质粒pHPV16为模板,采用PCR技术扩增HPV16主要衣壳蛋白L1(1-471aa)的基因序列,同时人工合成E7蛋白CTL表位E749-57的编码序列,将两个目的基因逐步连接构建重组质粒pUC19HPV16L1/E7CTL,HPV16L1/E7CTL片段经质粒pFastBac1转位至BacmidDNA后通过PCR鉴定重组杆粒。结果成功构建了包含融合基因HPV16L1/E7CTL片段的克隆质粒puc19HPV16L1/E7CTL,制备了L1/E7CTL重组杆状病毒载体杆粒BACMIDDNA。结论采用分子克隆技术构建了包含L1/E7CTL的重组杆粒BACMIDDNA,有利于下一步融合蛋白的表达,制备一种能够更有效激发CTL杀伤作用的嵌合病毒样颗粒疫苗,为疫苗研制打下基础。
简介:目的:探讨男性不育症患者精子DNA完整性与精子常规参数及形态的相关性。方法:纳入2016年1月至12月于我院就诊的男性不育症患者72例为观察组,同期体检健康男性100例为对照组。采用WLJY-9000型彩色精子质量检测系统检测精液常规参数及精子形态学参数,采用精子染色质扩散法(SCD)分析精子DNA完整性。对比观察组与对照组各检测指标差异。将观察组进一步划分为少精症组(n=16)、弱精症组(n=33)、白细胞精子症组(n=23),对比各小组检测指标的差异。分析观察组中精子DNA完整性与其它指标的相关性。结果:观察组精子浓度、精子存活率、前向运动精子占比、正常形态精子占比、精子DNA完整性均明显低于对照组,差异有统计学意义(P〈0.05)。少精症组精子浓度最低,弱精症组精子存活率最低,白细胞精子症组前向运动精子占比及正常形态精子占比最低,上述差异均有统计学意义(P〈0.05)。观察组精子DNA完整性与精子浓度、精子存活率、前向运动精子占比、正常形态精子占比均呈正相关(r=0.398、0.304、0.662、0.404,P均〈0.01)。结论:不孕症男性精子DNA完整性、精子浓度、精子存活率、前向运动精子占比、正常形态精子占比均明显下降,且精子DNA完整性与后4项观察指标均呈明显正相关。
简介:目的:探讨TCT,HC2-HPV-DNA检测和阴道镜检查在宫颈癌及癌前病变筛查中的联合应用价值.方法:1236例女性作为研究对象,依次进行HC2-HPV-DNA检测、TCT检查、阴道镜检查和病理学检查.结果:HC2-HPV-DNA检测,TCT和阴道镜检查的阳性率均显著小于病理学检查阳性率(x2=40.084,P<0.01;x2=45.008,P<0.01;x2=50.344,P<0.01),上述三项检查中,均为阳性者19例(1.5%),任意两项阳性者35例(2.8%),任意一项阳性者78例(6.3%).结论:TCT,HC2-HPV-DNA检测和阴道镜检查均有一定的局限性,但上述方法联合应用将大大提高筛出率,为患者及时治疗赢得宝贵时间.