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: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: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:目的评价抗苍白螺旋体IgM抗体检测对梅毒的临床意义。方法用酶联免疫吸附测定法对72例梅毒患者进行特异性IgM抗体检测,并与RPR、TPPA结果进行比较分析。结果血清抗苍白螺旋体IgM抗体在一期梅毒的阳性率为73.3%(11/15),在二期梅毒的阳性率为88.9%(16/18),二者比较无显著性差异(x~2=1.6363,P>0.10)。在潜伏梅毒,IgM抗体阳性率为26.1%(6/23),显著低于早期显性梅毒(x~2=17.6189,P<0.005)。在一期、二期和潜伏梅毒,RPR和TPPA的阳性率均为100%。入组前2-24个月己经给予正规抗梅治疗的梅毒16例,其中IgM抗体阳性2例。结论在本研究中,检测特异性IgM抗体诊断一期梅毒并下优于RPR和TPPA。IgM抗体在潜伏梅毒敏感性低,其诊断应依靠RPR和TPPA。目前不推荐单独检测抗梅毒IgM抗体来监测病情和判断疗效。抗苍白螺旋体IgM抗体的临床意义有待更深入的研究。
简介:目的通过对梅毒螺旋体(treponemapallidum,TP)脂肽激活巨噬细胞产生细胞因子以及脂肽所诱导的免疫耐受对信号通路的影响进行研究,为进一步完善TP感染人体的免疫学过程,解释TP感染人体后引起的血清固定现象提供理论依据。方法不同浓度的TP脂肽刺激经PMA诱导THP-1细胞转化的巨噬细胞,利用酶联免疫吸附试验(ELISA)检测其分泌细胞因子的水平,并通过阻断CD14受体后,检测巨噬细胞对合成脂肽的反应能力以及合成脂肽耐受刺激后诱导巨噬细胞产生免疫耐受的能力。采用Westernblot法检测细胞内的信号转导分子。结果3种合成脂肽诱导巨噬细胞分泌白细胞介素(IL)-β及IL-8的能力随着脂肽浓度升高而递增。CD14受体阻断后,IL-1β及IL-8分泌水平均明显减少。合成脂肽经耐受后刺激的IL-1β及IL-8分泌水平明显低于直接刺激的细胞因子的水平。合成脂肽在耐受刺激下,其巨噬细胞内的信号转导分子toll样受体2(tolllikereceptor2,TLR2)和核转录因子kappaB(NF-κB)P65的蛋白表达显著减少。结论3种合成脂肽均能诱导巨噬细胞产生IL-1β及IL-8。合成脂肽通过激活巨噬细胞膜表面CD14受体分子,活化下游信号通路,从而诱导细胞因子产生。合成脂肽能够诱导巨噬细胞模型产生免疫耐受,其可能机制为通过TLR2激活下游NF-κB信号通路,减少炎性细胞因子产生。
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.
简介:尖锐湿疣(Condylomaacumiatum,CA)又称性病疣(anogenitalwarts),是由人乳头病毒引起的,其发病率在中国仅次于淋病,居性传播疾病,第二位。在极少数病人可恶变诱发鳞癌对CA进一步的超微结构进行观察,探讨这些超微结病变在本病发病机理上的作用是十分必要的。更多还原