第8章微生物感染的病原学检查法

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1、MedicalMicrobiologyDepartmentofMicrobiology,HMU第一篇微生物学基础第8章微生物感染的病原学检查法LaboratoryDiagnosisZhao-HuaZhong,Prof.教学大纲掌握内容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

2、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OutlineLaboratorydiagnosisforbacterialinfectionsMorphological--microscopicMolecularSerologicalLaboratorydiagnosisforviralinfectionsMorphological--electronmicroscopicIsolationMolecular:nucleicaci

3、dsandproteinsSerologicalLaboratorydiagnosisforfungalinfectionsMorphological--microscopicIsolationAssaysMorphologicalassaysLightorelectronmicroscopiesIsolationanddifferentiationSerologicalassaysAntigen-antibodyassaysMolecularassaysMicroorganism’sgene(DNA&RNA)PrinciplesforSpecime

4、nCollectionSterilemanipulation,avoidcontaminationObtainaspecimenfromtheinfectedsiteaccordingtothediseasephasebeforeadministratingantibioticsDoubleseraforantibodydetections,acuteandrecoveryphaseeachPositiveifthetiterofthelaterspecimenisabove4XtothefirstspecimenTransport,andstore

5、correctlyassoonaspossibleStoreat-4oCformostbacterialspecimens,butkeepwarmforNeisseriameningitidisandNeisseriagonorrhoeaeStoreat-4oC~-70oCforviralspecimensLaboratoryDiagnosisforBacterialInfectionsMorphologicalProteinandNucleicacidsAntibodyBacteriologicalDiagnosisIdentifyingtheor

6、ganismMorphologicalIsolation&cultureBiochemicalreactionSerologicalassaysPathogenesis&antibioticssusceptibilityAnimalexperimentVirulencetestantibioticssusceptibilityMorphologicalNon-stainedmicroscopicobservationDark-fieldmicroscopyObservingthemovementoflivebacteriaStainedmicrosc

7、opicobservationsGramstainAcid-faststainFluorescencestainIsolation&Culture:ColonySizeShapeColorSurfacefeaturesSmooth-RoughTransparencyHemolysisBiochemicalReactionsSugarFermentationH2STestCitrateutilizationSerologicalAssaysDetectionantibodyinthepatient’sserumAcurrentinfectionshou

8、ldbeIgMpositiveA4-foldorgreaterriseonantibodytiterbetw