2022年医学专题—病原学检测法.ppt

第一篇微生物学(wēishēnɡwùxué)基础第8章微生物感染(gǎnrǎn)的病原学检查法LaboratoryDiagnosis第一页，共六十二页。(jiàoxuédàɡānɡ)掌握内容(nèiróng)细菌•细菌感染的微生物学检查程序•直接涂片镜检、分离培养和鉴定、生化试验、血清学试验及动物实验•病原菌抗原成分、核酸及其他成分的检测方法•血清学诊断原则及常用的方法病毒•病毒分离鉴定方法及病毒在培养细胞中的增殖的指标•病毒数量与感染性的测定（TCID50、PFU的原理、应用）•病毒成份的检测（抗原、核酸检测）•病毒抗体检测常用方法的原理及应用、双份血清诊断意义熟悉内容标本的采集与送检原则革兰染色和抗酸染色的原理细菌药敏试验真菌感染的微生物学检查法了解内容组织培养；聚合酶链反应、蛋白印迹技术的原理和应用第二页，共六十二页。LaboratorydiagnosisforbacterialinfectionsMorphological--microscopicMolecularSerologicalLaboratorydiagnosisforviralinfectionsMorphological--electronmicroscopicIsolationMolecular:nucleicacidsandproteinsSerologicalLaboratorydiagnosisforfungalinfectionsMorphological--microscopicIsolation第三页，共六十二页。第四页，共六十二页。MorphologicalassaysLightorelectronmicroscopiesIsolationanddifferentiationSerologicalassaysAntigen-antibodyassaysMolecularassaysMicroorganism’sgene(DNA&RNA)第五页，共六十二页。PrinciplesforSpecimenCollectionSterilemanipulation,avoidcontaminationObtainaspecimenfromtheinfectedsiteaccordingtothediseasephasebeforeadministratingantibioticsDoubleseraforantibodydetections,acuteandrecoveryphaseeachPositiveifthetiterofthelaterspecimenisabove4XtothefirstspecimenTransport,andstorecorrectlyassoonaspossibleStoreat-4oCformostbacterialspecimens,butkeepwarmforNeisseriameningitidisandNeisseriagonorrhoeaeStoreat-4oC~-70oCforviralspecimens第六页，共六十二页。LaboratoryDiagnosisforBacterialInfectionsMorphologicalProteinandNucleicacidsAntibody第七页，共六十二页。IdentifyingthePathogenesis&organismantibioticssusceptibilityMorphologicalAnimalexperimentIsolation&cultureBiochemicalreactionVirulencetestSerologicalassaysantibioticssusceptibility第八页，共六十二页。Non-stainedmicroscopicobservationDark-fieldmicroscopyObservingthemovementoflivebacteriaStainedmicroscopicobservationsGramstainAcid-faststainFluorescencestain第九页，共六十二页。第十页，共六十二页。第十一页，共六十二页。第十二页，共六十二页。第十三页，共六十二页。第十四页，共六十二页。第十五页，共六十二页。SizeShapeColorSurfacefeaturesSmooth-RoughTransparencyHemolysis第十六页，共六十二页。SugarFermentationH2STestCitrateutilization第十七页，共六十二页。Detectionantibodyinthepatient’sserumAcurrentinfectionshouldbeIgMpositiveA4-foldorgreaterriseonantibodytiterbetweentheacuteserumsampleandtheconvalescentserumsampleMajordrawbacksAsingleIgGantibodytiterisdifficulttointerpretbecauseitisunclearwhetheritrepresentsacurrentorapreviousinfectiontheconvalescentsampleisusuallytaken10-14daysaftertheacutesample.Bythistime,thepatienthasoftenrecoveredandthediagnosisbecomesaretrospectiveoneSomeexceptionsIncertaindiseases,asingletiterofsufficientmagnitudecanbeusedaspresumptiveevidenceofacurrentinfection第十八页，共六十二页。AnimalsMouseGuineaPigRabbitDogMonkeyInoculationroutesSubcutaneousIntradermalIntraperitonealIntravenousIntracranial/intracerebralIntraspinalIntranasalLavage第十九页，共六十二页。Medianlethaldose,LD50Medianinfectivedose,ID50ElekPlateDiphtheroto...