感染病学基础-病原学诊断-8-30.pptx

病原学诊断

Diagnosticmicrobiology;内容

Contents

;;〔1〕目的：直接检测微生物〔细菌、真菌、寄生虫虫卵幼虫和成虫、感染的细胞中的病毒包涵体〕;初步或明确鉴定微生物;;;;;;;〔3〕检查方法：;〔1〕成功取决于：

微生物的生物学特性

感染的部位

患者对感染的免疫反响

适宜培养基〔三种类型〕和条件;培养基类型〔细菌和真菌〕;培养基类型〔衣原体、立克次体和全部病毒〕：细胞培养;〔1〕分子种类：

DNA

RNA

蛋白质

〔2〕优势：

适宜于不能别离或免疫学检查的微生物检查；

检测基因型或突变〔eg.耐药〕

敏感、特异和平安；;Restrictionfragmentlengthpolymorphism〔RFLP，限制性片段长度多态性〕distinctionofDNAfrombacterialstrainsseparatedbypulsed-fieldgelelectrophoresis.

Lanes1to3showSma1restrictionendonuclease-digestedDNAfrombacteriafromtwofamilymemberswithnecrotizingfasciitisandfromtheirphysician(pharyngitis).Lanes4to6arefromunrelatedStreptococcuspyogenesstrains;DNAprobe〔DNA探针〕analysisofvirus-infectedcells.SuchcellscanbelocalizedinhistologicallypreparedtissuesectionsusingDNAprobesconsistingofasfewasninenucleotidesorbacterialplasmidscontainingtheviralgenome.AtaggedDNAprobeisaddedtothesample.Theappropriatesubstrateisaddedtocolor

thenucleiofvirallyinfectedcells.;Insitulocalization〔原位杂交〕ofcytomegalovirus(CMV)infectionusingageneticprobe.CMVinfectionoftherenaltubulesofakidneyislocalizedwithabiotin-labeled,CMV-specificDNAprobeandisvisualizedbymeansofthehorseradishperoxidase-conjugatedavidinconversionofsubstrate,inamannersimilartoenzymeimmunoassay.;多聚酶链反响;DNAchip〔基因芯片〕

16srRNA测序或全基因组测序

蛋白组和质谱;检测和定量抗原

评价抗体反响

评估是否暴露或免疫接种;Analysisofantigensandantibodiesbyimmunoprecipitation〔免疫沉淀〕;Immunofluorescenceandenzymeimmunoassaysforantigenlocalizationincells.Antigencanbedetectedbydirectassaywithantiviralantibodymodifiedcovalentlywithafluorescentorenzymeprobe,orbyindirectassayusingantiviralantibodyandchemicallymodifiedantiimmunoglobulin.Theenzymeconvertssubstratetoaprecipitate,chromophore,orlight.;Immunofluorescencelocalizationofherpessimplexvirus–infectednervecellsinabrainsectionfromapatientwithherpesencephalitis.;Flowcytometry