简介:Objective:Infectionofhumanpapillomavirusincondylomaacuminatum(CA)wasdetectedbyrealtimefluorescencequantitativePCR(FQ-PCR)technique.Methods:SpecimensofCA-DNAquantificationfrom94caseswereexaminedbyrealtimeFQ-PCRtechniqueand32caseswerecomparedwiththesamemethodafter10-daystreatment.Results:CA-DNAwasfoundinallpatients,withanaverageof4.0×10^6copies/ul.After10daysoftreatment,theaveragewas2.1×10^5copies/ul.TherewasasignificantdifferenceintheaverageamountofCA-DNAbeforeandafterthetreatment.Conclusion:RealtimeFQ-PCRisagoodmethodforexaminingCA-DNAamountanditcandirectthetreatmentofCA.
简介:Objective:Tocomparethesensitivityandspecificityofthecervical/urethralswabswithvoidedurinespecimensforthedetectionofgenitourinarytractinfectionwithChlamydiatrachomatisanddeterminewhetherurinespecimenscanreplacethecervical/urethralswabsindetectionofC.trachomatis.Methods:Thematchedcervical/urethralswabsandvoidedurinespecimenswerecollectedfrom569patientsofSTDclinics.Polymerasechainreaction(PCR)assayspecificforC.trachomatisplasmidDNAandrapidantigentesting(Clearviewassay)wasusedtodetectC.trachomatis.Standardcriteriathatdefined""""true""""positiveincluded:1)positivePCRresultsbothincervical/urethralswabandvoidedurinespecimenor2)positivevoidedurineresultsbothbyPCRassayandclearviewtestor3)positiveresultsinbothPCRassayofcervical/urethralswabandclearviewtestofvoidedurine.Forstatisticalanalysis,thechi-squaretestwasused.Results:TheprevalenceofC.trachomatisinpatientswithsymptomswas12.1%(28/231)inwomenand10.4%(10/96)inmen,withnosignificantdifferencebetweenthem(x^2=0.21,P>0.05).TheprevalenceofC.trachomatisinpatientswithnosymptomswas11.0%(11/100)inwomenand15.5%(22/142)inmen,withasignificantdifferenceexistingbetweenthem.(x^2=4.0,P<0.05).Nosignificantdifference(P>0.05)existedbetweenPCRtestingofswabs(sensitivity87.3%;specificity99.2%)andPCRtestingofurine(sensitivity88.7%;specificity98.8%).Asforclearviewassay,sensitivitywas60.6%andspecificitywas100%.Conclusions:PCRassayissuperiortoclearviewindetectingC.trachomatis.AlthoughbothPCRtestingofswabsandPCRtestingofurinespecimensbothhavehighsensitivityandspecificity,urinespecimentestingismorecost-effective,practicalandnoninvasive.ThusurinespecimenscantaketheplaceoftheswabsinPCRtestingforchlamydia.
简介:目的评价PCR检测女性自取阴道下端拭子标本诊断淋病的检验效果方法每例入选者均取2个宫颈拭子标本和1个自取阴道下端拭子标本,1个宫颈拭子标本和自取阴道下端拭子标本行淋球菌PCR检测,另1个宫颈拭子标本行淋球菌培养。结果共有298例入选者。诊断出淋病31例。宫颈拭子培养、宫颈拭子PCR、自取阴道下端拭子标本PCR的敏感性分别是75.8%(25/33),87.9%(29/33)和97.0%(32/33),特异性分别是100%(265/265),99.6%(264/265)和99.6%(264/265)。结论自取阴道下端拭子标本PCR检测淋球菌的敏感性至少不低于传统的宫颈拭子,自取阴道下端拭子标本可以替代宫颈拭子行PCR诊断女性淋病。
简介:Abstract:TheP24antigentest,HIVRNAPCRtest,HIVisolation/cultureandfourth-generationHIVuniformAg/AbassayarebeingutilizedindiagnosingacuteHIVinfectionindifferentlabs.ManyfactorslimittheuseofscreeningforacuteHIVinhigh-riskpopulations,inblooddonorsandduringvoluntaryHIVtesting,including,cost,technique,sensitivityandspecificity.InthisreviewweexploreanewNAATmethodwhichinvolvesHIVRNART-PCRonpooledsamples.Thistechniqueisabletoscreenforacuteinfectionsinalargetestingvolumeandmayheusedasascreeningmethodinhigh-riskpopulationsandblooddonors.
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:建立一种快速、稳定、灵敏且可定量的检测马尔尼菲青霉病的SYBRGreenⅠ实时荧光定量PCR方法。方法:根据马尔尼菲青霉ITS序列,设计并合成引物,通过常规PCR扩增目标序列后,将鉴定正确的目的基因片段克隆入pGEM-T载体中,转化大肠杆菌JM109,经PCR及测序鉴定后得到阳性重组质粒,作为标准品模板建立SYBRGreenⅠ荧光定量PCR标准曲线和溶解曲线,得出拷贝数与循环阈值之间的线性关系曲线表达式,并做灵敏性、特异性试验。结果:荧光定量PCR标准曲线阈值与模板浓度呈良好的线性关系,溶解曲线特异,相关系数为1.938,最低检测的拷贝数为8.5个/反应管。结论:建立了检测马尔尼菲青霉病的SYBRGreenⅠ实时荧光定量PCR方法,为该病的早期诊断及定量分析感染程度及对治疗的指示作用奠定了良好的基础。
简介:Objectives:TodevelopamethodofsimultaneousPCRdetectionofHaemophilusducreyi,Treponemapallidum,andHerpesSimplexVirusTypes1and2fromgenitalulcersamongpatientsattendingSTDclinicsinGuangzhou,China;andevaluatetheclinicalapplicationofmultiplexPCR(M-PCR)assayfordiagnosingtheetiologyofgenitalulcerdiseases(GUD).Methods:244patientswithagenitalulcerwereevaluated.ClinicaletiologyofGUDwasbasedonphysicalappearanceandmicrobiologicevaluationsthatincludeddarkfieldmicroscopyexamination(D-F)andserologytestforsyphilis(STS).SwabsofeachgenitalulcerweretestedforHSVantigenbyenzymeimmunoassay(EIA)andprocessedinanM-PCRassayforsimultaneousdetectionofT.pallidum,HSVandH.ducreyi.Results:ThestandardstrainsofT.pallidum,HSVandH.ducreyiwereamplifiedbyM-PCR,producingamplifiedproductsof260bp,432bp,170bp,respectively.ThesensitivityofM-PCRis102pgDNA.M-PCRassayforT.pallidum,HSVandH.ducreyishowedgoodagreementwhencomparedwithD-FdetectionforT.pallidum,STS,H.ducreyicultureandEIAforHSVantigen(Kappascoresare0.774,0.704,0.793,0.756,respectively).Conclusions:TheM-PCRisaconvenient,accurateandreliableassayforthedetectionofT.pallidum,HSVandH.ducreyifromgenitalulcers,andcanbeusedasamethodofdiagnosingtheetiologyofGUD.
简介:目的比较荧光多重PCR和血清型特异性抗体测定在生殖器疱疹临床诊断中的应用价值及评价各自的优缺点。方法以细胞培养法作为“金标准”对照,分别用荧光多重PCR和血清型特异性抗体检测法对121例临床诊断为生殖器疱疹的标本进行检测。结果以培养法作标准,并通过结果的差异性分析,荧光多重PCR的敏感性为100%,特异性为88.89%;血清型特异性抗体测定则分别为77.68%和77.78%,荧光多重PCR的敏感性和特异性均显著高于血清型特异性抗体测(P<0.05),但前者不能检测出无皮损患者的HSVDNA,而后者可检测出无皮损患者中的HSV抗体。结论荧光多重PCR和血清型特异性抗体检测各有其自身的优缺点,单独用PCR和其它病毒分离的方法或单独使用血清特异性抗体检测的方法来诊断生殖器疱疹都是不完整的,均可造成漏诊。临床上将两种方法有机的结合起来应用能发挥各自的优势,取长补短,对早期、准确、快速地诊断生殖器疱疹及进行流行病学调查将有十分重要的意义和使用价值。
简介:目的:探讨腹腔镜下切除和开窗两种术式治疗输卵管壶腹部妊娠的效果,并进行对比分析。方法:选取我院妇产科2013年1月至2014年1月收治的90例输卵管壶腹部妊娠患者,随机将患者分成切除组和开窗组,每组各45例,在腹腔镜下分别行输卵管切除术和输卵管开窗术,分别观察两组患者围术期情况、术后T-HCG情况和术后再次妊娠情况。结果:两组患者在围术期手术时间、术中出血量等方面并无统计学意义,P〉0.05;术前血T-HCG和术后3d血T-HCG的下降比率比较无统计学意义,而术后1周血T-HCG下降比率和术后1个月血T-HCG下降至正常例数的比较具有统计学意义,P〈0.05;两组术后宫内妊娠、异位妊娠和继发不孕的比较没有统计学意义,P〉0.05。结论:腹腔镜下输卵管切除术与开窗术具有相同的治疗价值,而输卵管开窗术不能提高宫内妊娠率。
简介:主要总结并探讨几种常见的丘疹鳞屑性皮肤病在反射式共聚焦显微镜(reflectanceconfocalmicroscopy,RCM)下的镜像特征以及RCM技术在丘疹鳞屑性皮肤病辅助诊断中的应用进展。其中寻常性银屑病的RCM研究较多且相对深入,已建立了观察银屑病不同病期及疗程的量化指标。玫瑰糠疹的RCM研究比较缺乏,可能与该病相对无特异性组织病理学表现和有限的RCM分辨率有关。扁平苔藓的RCM诊断可以达到类似于特异性组织病理学改变的水准,但毛发红糠疹应用RCM观察其特异性组织病理表现相对困难。硬化性萎缩性苔藓和光泽苔藓的RCM研究虽少但其表现较为特异。RCM技术已为皮肤病的诊断和鉴别诊断提供了极大帮助,有待设备技术的更新改良和更多简便实用的应用研究来进一步提高其应用效果及前景。
简介:目的:比较子宫肌瘤不同手术方式对绝经过渡期女性患者卵巢功能、性功能和生活质量的影响。方法:回顾性分析2008年1月至2012年1月于我院确诊的60例子宫肌瘤患者资料,其中30例患者行子宫全切除术(I组),另外30例行子宫肌瘤剜除术(Ⅱ组),术后随访24个月。对患者两种手术术后卵巢功能、性生活质量和生活质量等指标进行分析。结果:两组手术均未出现死亡及严重并发症,Ⅱ组患者FSH、LH显著低于I组患者(t=4.376和4.112;P=0.018和0.001),E2显著高于后者,差异均具有统计学差异(t=34.981,P=0.021);Ⅱ组患者性功能和生活质量评分显著高于I组,差异均具有统计学意义(t=2.785和4.714;P=0.017和0.021)。结论:与子宫全切除术相比,子宫肌瘤剜除术对患者卵巢功能、性功能影响较小,患者生活质量更高,值得临床合理运用。