简介:ToestablisharapididentificationmethodforcommonpathogenicbacteriaonthebasisofmolecularbiologyandtoconstructapreliminaryPolymeraseChainReaction-CapillaryElectrophoresis-RestrictionFragmentLengthPolymorphism(PCR-CE-RFLP)databaseofbacteriaisolatedfromclinicalspecimensfrequently,183strainscollectedfromclinicalsamplesbelongingto12generaand19specieswhosebiochemicalcharacterizationscorrespondedtothetypicaloneswereexamined.ThegenomicDNAswereamplifiedbytwopairsoffluorescencelabeledprimersaimingat16SrRNAgeneand16S-23SrRNAspacerregiongenerespectivelyatthesametime.PCRproductswerethendigestedbyrestrictionendonucleaseHaeⅢin-completelybeforetakingcapillaryelectrophoresis.TheresultswiththePCR-CE-RFLPpatternsof16SrRNAgeneswerejustalikewithinsomegenera,butwhenitcomesto16S-23SrRNAspacerregiongenes,eachbacteriumshowedauniquepattern,whichcanbedistinguishedfromeachothereasily.ItseemsthatPCR-CE-RFLPpatternsof16SrRNAgenecouldonlybeusedtoclassifythebacteriaintofamilylevel,whereasthedataof16S-23SrRNAspacerregiongenecouldbeutilizedtoidentifythewholemicroorganismsaspreciselyasthespecieslevel.Inspiteofthedataofthespacerregiongenealonecanbesufficientlytoverifythewholebacteria,weinsistthatthe16SrRNAgenecouldbeofsomeassistantincasethatthereshouldbelotsoffamiliesofbacteria,inwhichsomesimilarones,withthesameRFLPdataof16S-23SrRNAspacerregiongene,maycoexist.ThisstudyprovesthattheutilityofPCR-CE-RFLPisaconvenient,rapidmethodtoidentifypathogenicbacteria,andisalsoaquickdiagnosismeasureforapplicationtoclinicaluse.
简介:【摘要】目的:分析荧光定量RT-PCR在流感病毒检测上的应用价值。方法:对23例确诊感染乙型流感病毒的咽拭子、相关机构提供的甲型流感病毒及乙型流感病毒进行检测分析,使用荧光定量RT-PCR检测,分析临床应用的价值及检测的准确率。结果:荧光定量RT-PCR检测乙型流感病毒呈阳性,且结果与其他病毒未见交叉情况。荧光定量RT-PCR检测不同浓度乙型流感病毒敏感度较高,检测的准确率是86.96%(20/23)。结论:荧光定量RT-PCR检测流感病毒的敏感度、准确性较高,可以作为疾病筛查的依据,建议推广应用。
简介:【摘要】目的 研讨轻中度痔疮采用自动痔疮套扎术治疗的效果。方法 选择70例轻中度痔疮患者,均来源于本院2019年3月-2019年11月期间收入,按治疗方法不同分成两组,对照组和观察组,组内分别有35例。对照组采取常规外剥内扎术治疗,观察组采取自动痔疮套扎术治疗,对比两组治疗效果和术后并发症发生情况。结果 观察组治疗总有效率为94.29%,显著高于对照组的77.14%(P<0.05);观察组术后并发症发生率为5.71%,显著低于对照组的22.86%(P<0.05)。 结论 轻中度痔病采用自动痔疮套扎术治疗能显著提高治疗效果,减少术后并发症发生。
简介:Thepurposeofthestudyistoestablishafluorescencequantitativereversetranscriptionpoly-merasechainresponse(FQ-RT-PCR)methodforthequantitativedeterminationofIL-2mRNAandIL-4mRNAinThcells,withwhichtheThcellsstatusofthepatientswithgynaecologicaltumorsandchronicrenalfailure(CRF)canbeanalyzed.IL-2cDNAandIL-4cDNAwereprepared,andtheplasmidpMD18carryingIL-2cDNAorIL-4cDNAfragmentwasconstructedandclonedasthetemplateforquantitativedetermination.Theprimersandprobeslabelledwith6-carboxy-fluorescein(FAM)and6-carboxy-tetrarnethylrhodamine(TAMRA)wereprepared,andtheexperimentalconditionswereoptimizedtosetuptheFQ-RT-PCRmethodforquantitativedeterminationofIL-2mRNAandEL-4mRNA.Thcellsenrichedfromperipheralbloodmononuclearcells(PBMCs)of20healthyvolunteers(HVs),16gynaecologicalbenign(GB)cases,18gynaecologicalmalignant(GM)tumorcasesand16chronicrenalfailure(CRF)patientsweretestedforIL-2mRNAandIL-4mRNAbyFQ-RT-PCR.Thehouse-keepinggeneβ-actinwasusedastheinternalcontrolgeneoftheexperiment.Thestandardcurveforlogconcentrationofseriesofquantitativetemplatesvsthresholdcycle(CT)wasestablishedbylinearregression,andthelinearrangewas102-107copies/μl.TheimprecisiontestshowedtheCVofinter-assayandintra-assayofahighcontentsamplebyFQ-RT-PCRwere7.8%and12.5%,respectively.TheCVofinter-assayandintra-assayofalowcontentsamplewere10.8%and19.5%,respectively.TheIL-2mRNAexpressionsinThofthepatientswithgynaecologicalmalignanttumor(comparedwiththeHVsandthepatientswithgynaecologicalbenigndisease)andinThoftheCRFpatients(comparedwiththeHVs)weredeclinedsignificantlyandatthesametimetheIL-4mRNAexpressionincreasedsignificantly(P<0.001).Asimple,sensitiveandaccurateFQ-RT-PCRmethodforthequantitativedetectionofIL-2mRNAandIL-4mRNAhasbeenestablished.TheIL-2mRNAandIL-4m
简介:【 摘 要 】 目的: 讨论 PCR 产物酶联免疫检测法在乳腺癌临床检验中的应用价值评估。 方法 : 选取我院治疗的乳腺癌的患者 50 例,均对患者使用 PCR 产物酶联免疫检测法检测,使用手术治疗,在手术后进行病理检测,以病理检测的结果为金标准。 结果 : PCR 产物酶联免疫检测 与手术病理检测结果相比,差别较小 (
简介:Theaimofthisstudywastoanalyzethepointmutationoftheexon1atcodon54ofthemannose(ormannan)-bindinglectin(MBL)geneinhealthyindividualsofChineseHansandMongolianpopulation,andtofindoutanyassociationbetweentheplasmalevelsofMBLandthegenemutationfrequencyinbethgroupsofindividuals.Bloodsampleswerecollectedrandomlyfrom56healthyindividualsofChineseHansand37Mongolian.ThedetectionofthepointmutationsoftheMBLgenewasperformedbypolymerasechainreaction-restrictionfragmentlengthpolymorphism(PCR-RFLP)anddetectionsforplasmalevelsofMBLweredeterminedbyusingMBLELISAkits.AMBLPCRmethodofassaywasestablishedwithhighspecificity,andgoodreproducibility.ByoptimizingthePCRcondition,theoptimalannealingtemperaturewas55℃,andthelowestdetec-tionlimitwas160pg.Nobandswerefoundinnon-specificitysamples(HAV,HBV,HCVandTB),andthesequencesofPCRproductswerethesameastheexpectedones.AlsoaMBLPCR-RFIPwasestablished.Uponelectrophoresisofthedigestedproductsin3%agarosegel,therewere3patterns:inwhich2bandscorrespondtomoleculeweight232bpand93bp;1band,correspondstomoleculeweight325bpand3bandscorrespondtomoleculeweight325bp,232bpand93bp,respectively.Threebandsof325bp,232bpand93bpofpointmutationswerefoundatcedon54ofMBLcedinggene.FrequenciesinhealthyHanandMongolianpopulationwere0.2321and0.1757respectively.TheaverageplasmaMBLconcentrationwas1998.750μg/L,withSDof1505.152in56healthyHanpopulationand2525.676μg/L,withSDof1955.188in37Mongolian.AnegativecorrelationbetweenMBLconcentrationandgenemutationfrequencywasfoundinhealthyHanpopulation.Frequencyofpointmutationwas1.00whentheMBLconcentrationswerebelow100μg/L;frequencyofpointmutationwas0.4524whentheconcentrationwas100μg/Lto1000μg/L;andthefrequencyofpointmutationwas0.0156whentheconcentrationwaso
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.
简介:【摘要】目的:探究分析自动痔疮套扎术治疗痔疮出血的效果和患者不良反应情况。方法:在2018年8月到2020年8月期间本院收治的痔疮出血患者中选择90例作为观察对象,根据治疗方式的不同将患者分为实验组和对照组两组,每组患者各45例。对照组患者给予传统痔疮切除术治疗,实验组患者给予自动痔疮套扎术治疗,对比两组患者治疗前后的VAS评分、不良反应情况和随访两个月后的复发情况。结果:实验组痔疮患者经过手术治疗后,和对照组患者相比,VAS评分明显下降,且术后疼痛、出血、切缘水肿等不良反应和随访复发率明显更低,差异具有统计学意义(p<0.05)。结论:自动痔疮套扎术用于治疗痔疮出血具有较好的临床疗效,能够有效缓解患者的疼痛感,明显改善患者的相关症状,促进患者身体康复,值得临床广泛应用。
简介:【摘要】目的:对行内镜下食管静脉曲张套扎术患者予以优质服务护理干预的临床效果进行分析评价。方法:对于本次研究,选取的研究对象均是于我院2018年6月-2019年7月收治的68例行内镜下食管静脉曲张套扎术患者,将所有患者随机分为两组,每组34例,对照组行内镜下食管静脉曲张套扎术患者予以常规护理,实验组行内镜下食管静脉曲张套扎术患者予以优质服务护理干预,对两组患者的护理结果进行统计分析。结果:经过护理,实验组食管静脉曲张患者的疼痛评分明显低于对照组患者,结果具有统计学意义(P<0.05);且实验组食管静脉曲张患者的护理满意度明显高于对照组,结果具有统计学差异(P<0.05)。结论:对行内镜下食管静脉曲张套扎术的患者予以优质服务护理干预,能够有效缓解患者的术后疼痛,提高患者的护理满意度,具有临床意义。