临床常见病原体检测英文版教学课件

临床常见病原体检测

Examinationofclinicalcommonpathogens呼吸内科熊维宁DepartmentofRespiratoryMedicineXiong,Weining临床常见病原体检测

Examinationofclin1目的

Objective确定感染的发生和性质，及早明确诊断；Determinetheoccuringandnatureofinfection,andmakediagnosisearly;尽早选择适当的治疗方案；Selecttheappropriatetreatmentassoonaspossible;采取有效的预防措施，防止感染可能广泛传播所造成的危害。Takeeffectivepreventionmeasurestopreventinfectiontocausewidespreaddamage.目的

Objective确定感染的发生和性质，及早明确诊断；2根本程序

BasicProcedures正确采集和运送标本；Correctcollectionandtransportationofspecimens初步诊断：直接镜检，免疫学和分子生物学检测；Initialdiagnosis:directmicroscopy,immunologyandmolecularbiologyexamination;确定诊断：病原体的别离、鉴定及药敏实验，报告结果；Determineddiagnosis:pathogenisolation,identificationandsensitivitytest,reportingresults;合理用药Appropriatedruguse根本程序

BasicProcedures正确采集和运送标本3第一节

SectionOne标本的采集运送、实验室评价和检查方法Transportationandcollectionofspecimen,laboratoryevaluationandexaminationmethods第一节

SectionOne标本的采集运送、实验室评价和检4一、标本采集和运送One.CollectionandtransportationofSpecimen一、标本采集和运送5根本原那么

Basicprinciples根据病史与临床表现确定标本采集的时间、部位、种类和数量，尽量采集病变明显部位标本；Accordingtohistoryandclinicalmanifestationstodeterminethecollectiontime,location,typeandquantityofspecimens,collectspecimensatsignificantpartsoflesions;无菌操作，防止污染；Useaseptictechniquetopreventcontamination;采取适宜的方式进展储存，尽快送检；Appropriatewaytostore,assoonaspossibleforexamination;要视所有标本为传染品，高度危险性的标本要有明显标识，急症或危重患者标本要特别注明。Toviewallthespecimensforinfectious,highriskspecimensshouldbeclearlymarked,acuteorcriticallyillpatientspecimensshouldbespecified.根本原那么

Basicprinciples根据病史与临床表6临床常见病原体检测英文版教学课件7临床常见病原体检测英文版教学课件8〔二〕尿液

(Two)Urine无菌采集中段尿；Sterilecollectionofmid-portionurine;如考虑厌氧菌感染，采取膀胱穿刺法采集标本，无菌厌氧小瓶运送；Ifconsiderationofanaerobicinfections,applybladderpuncturetocollectspecimens,transportthemwithsterileanaerobicvial;排尿困难者考虑导尿采集标本。Applycatheterizationtocollectspecimensfordysuriapatients.〔二〕尿液

(Two)Urine无菌采集中段尿；9〔三〕粪便

(Three)Stool挑取脓、血或粘液局部于清洁容器中送检；Pickpus,bloodormucusfromstollintocleancontainersforexamination;排便困难者或婴儿采用直肠拭子采集，置于有保存液的试管内送检；Applyrectalswabforinfantsordifficultdefecation,placeitwithpreservationsolutioninthetesttubeforexamination;疑心霍乱弧菌感染引起的腹泻，将标本置于碱性蛋白胨水或卡-布〔Cary-Blair〕运送培养液送检；SuspectedinfectionscausedbyVibriocholerae,thespecimenswereplacedinalkalinepeptonewaterorcard-cloth(Cary-Blair)transportmediumforexamination;传染性腹泻应连续送检3次。Continuousexamination3timesforinfectiousdiarrhea.〔三〕粪便

(Three)Stool挑取脓、血或粘液局部于10〔四〕呼吸道标本

(Four)Respiratoryspecimens类型：鼻咽拭子，痰和经气管采集的标本；Types:nasopharyngealswab,sputum,andspecimenscollectedthroughtracheal上呼吸道存在正常菌群，在采集标本与结果分析时应予考虑。Thereisnormalflorainupperrespiratorytract,thespecimenscollectionandresultsanalysisshouldbeconsidered.〔四〕呼吸道标本

(Four)Respiratorysp11〔五〕脑脊液与其他无菌体液

(Five)Cerebrospinalfluidandothersterilebodyfluids脑脊液应立即保温送检或床边接种；CSFshouldbetransportedimmediatelywithinsulationorculturedbedside；胸腔积液、腹腔积液和心包积液等应采集较大量标本送检，离心后再接种培养。Pleuraleffusion,ascitesandpericardialeffusionwithalargevolumeshouldbecollected,centrifugatedandthencultured.〔五〕脑脊液与其他无菌体液

(Five)Cerebrosp12〔六〕眼、耳部标本

(Six)Eye,earspecimens拭子采样Swabsamples〔六〕眼、耳部标本

(Six)Eye,earspeci13〔七〕泌尿生殖道标本

(Seven)Urogenitalspecimens男性：无菌采集尿道口分泌物或前列腺液；Male:urethraorprostaticfluidsecretionswithsterilecollection;女性：无菌采集阴道或宫颈分泌物。Female:vaginalorcervicalsecretionscollectedaseptically.〔七〕泌尿生殖道标本

(Seven)Urogenital14〔八〕创伤、组织和脓肿标本

(Eight)Trauma,organization,andabscessspecimens对损伤范围较大的创伤，从不同部位采集多份标本；Onawiderrangeoftraumainjuries,specimenscollectedfrommanydifferentparts;采集部位应首先去除污物、消毒皮肤；Firstlyremovedirt,disinfecttheskinaroundcollectionsite;标本较少那么需参加无菌生理盐水以防枯燥；Fewersamplesmustbeaddedtosterilesalinetopreventdrying;开放性脓肿及脓性分泌物：用无菌棉纤采取脓液或病灶深局部泌物；Openabscessandpurulentsecretions:collectdeepdischargepusorlesionswithsterilecottonfibers;封闭性脓肿用注射器抽取；Collectspecimeninclosedabscesswithasyringe;疑心厌氧菌感染，应隔绝空气采集。Suspectedanaerobicinfections,collectionshouldbeisolatedfromair.〔八〕创伤、组织和脓肿标本

(Eight)Trauma,15〔九〕血清

(Nine)Serum用于检测特异性抗体；Forthedetectionofspecificantibodies;血液自然凝固后吸取血清，灭活补体。Drawserumafternaturalcoagulationofblood,theninactivatecomplement.〔九〕血清

(Nine)Serum用于检测特异性抗体；16二、标本的实验室质量评估标准

Two.Standardoflaboratoryqualityassessmentofspecimens二、标本的实验室质量评估标准

Two.Standardo17检验申请单的根本内容

ThebasiccontentofanapplicationforexaminationThepatient,snameandhospitalnumber.Ageandsex.Collectiondateandtime.Suspecteddiagnosis.Exactnatureandsourceofthespecimen.Immunizationhistoryandantimicrobialtherapy.Objective.Signedbyphysician.检验申请单的根本内容

Thebasiccontento18标本接收和拒收准那么

Receptionandrejectioncriteriaforspecimens1Theinformationonthelabeldoesnotexistornotmatchtheinformationontherequisition;2Delayindeliveryofthespecimens;3Thespecimenisleaking;4Thespecimenhasbeentransportedattheimpropertemperatureorinimpropermedium.标本接收和拒收准那么

Receptionandrejec195Contaminatedsamples;6Thequantityofspecimenisinsufficientfortesting;7Duplicatesubmissionofspecimens(exceptbloodcultures);8Thetransportationofspecimensofsevereinfectiousdiseasesshouldcomplywiththerelevantrules.5Contaminatedsamples;20三、检查方法

Three.Examinationmethods

三、检查方法

Three.Examinationmeth21〔一〕直接显微镜检查

(One)Directmicroscopicexamination不染色标本检查法：用于观察病原体的生长、形态与运动等特性；Notstainedspecimentestmethod:toobservethegrowth,shapeandmotioncharacteristicsofpathogens;评价：局部病原体可借此初步诊断。Evaluation:forsomepathogens,theinitialdiagnosiscanbetaken.〔一〕直接显微镜检查

(One)Directmicros22染色标本检查法：观察细菌的形态、染色性或观察宿主细胞内包涵体的特征；Stainedspecimentestmethod:toobservebacterialmorphology,stainingorobservethecharacteristicsoftheinclusionbodiesinhostcell;评价：为临床初步诊断提供依据。Evaluation:availablebasisforthepreliminaryclinicaldiagnosis.染色标本检查法：观察细菌的形态、染色性或观察宿主细胞内包涵体23革兰氏〔染色〕阳性球菌Gram(staining)-positivecocci革兰氏〔染色〕阴性杆菌Gram(staining)-negativebacilli革兰氏〔染色〕阳性球菌革兰氏〔染色〕阴性杆菌24抗酸染色阳性杆菌〔抗酸杆菌〕Acid-faststain-positivebacilli(Acid-fastbacilli)抗酸染色阳性杆菌〔抗酸杆菌〕25〔二〕病原体特异性抗原检查

(Two)Pathogen-specificantigentest免疫荧光技术Immunofluorescencetechnique酶联免疫技术ELISA化学发光技术Chemiluminescencetechnology乳胶凝集试验Latexagglutinationtest对流免疫电泳Counterimmunoelectrophoresis蛋白质芯片Proteinchips〔二〕病原体特异性抗原检查

(Two)Pathogen-s26病原体特异性抗原检测结果评价

Evaluationofpathogen-specificantigentestresults如果能排除穿插抗原的影响，病原体抗原检测可明确感染的病原体。Iftheimpactofcross-antigencanbeexcluded,pathogenantigentestcouldcleartheinfectionofpathogens.病原体特异性抗原检测结果评价

Evaluationofp27〔三〕病原体核酸检查

(Three)Pathogennucleicacidtest聚合酶链式反响：扩增病原体微生物特异的DNA或RNA片段；Polymerasechainreaction(PCR):amplifythespecificfragmentsofDNAorRNAofmicrobialpathogens;实时荧光定量PCR技术；Real-timePCR;核酸探针杂交技术：通过序列的探针与标本中的病原体的核苷酸杂交，用以了解病原体的有无；Nucleicacidprobehybridization:hybridizationbetweentheknownsequenceoftheprobeandthenucleotideofpathogensinthesamples,toexplorewhetherpathogensexist;基因芯片技术。GenechiporDNAmicroarray.〔三〕病原体核酸检查

(Three)Pathogennu28病原体核酸检查结果评价

EvaluationofpathogenDNAtestresults是检测病原体微生物最灵敏的方法，但具有一定的假阳性与假阴性；isthemostsensitivedetectionmethodformicrobialpathogens,buthassomefalsepositiveandfalsenegative;阳性只说明存在某种病原体的核酸，是否正被感染应结合临床具体分析。istheonlyshowthattheexistenceofapositivepathogennucleicacid,whetherbeinginfectedshouldbecombinedwithclinicalspecificanalysis.病原体核酸检查结果评价

Evaluationofpath29〔四〕病原体的别离培养与鉴定

(Four)Isolation,cultureandidentificationofpathogens1细菌感染性疾病病原体的别离培养1Isolationandculturethepathogensofbacterialinfections明确感染病原体；Clearthepathogens;为临床提供体外抗微生物药物敏感试验结果。Providevitrotestresultsofanti-microbialdrugsensitivityforclinical.〔四〕病原体的别离培养与鉴定

(Four)Isolatio302不能人工培养的病原体感染性疾病2Notcultivatedpathogensofinfectiousdisease将标本接种易感动物、鸡胚或行细胞培养。Specimenswereinoculatedintosusceptibleanimals,eggsorcelllines.2不能人工培养的病原体感染性疾病31〔五〕血清学实验

(Five)Serologicaltest特异性IgM可作为感染性疾病的早期诊断指标，且可区分原发与复发感染；SpecificIgMcanbeindicatorofearlydiagnosisforinfectiousdiseasesandcandistinguishbetweenprimaryandrecurrentinfections;特异性IgG，尤其双份血清的滴度呈4倍或4倍以上升高，考虑现症感染。ForIgG,thetiterofdoubleserumhigher4timesormorethan4times,currentpathogeninfectionshouldbeconsidered.〔五〕血清学实验

(Five)Serologicalte32第二节病原体耐药性检测

SectiontwoDetectionofpathogendrugresistance

第二节病原体耐药性检测

SectiontwoDetec33抗生素压力Antibioticpressure抗生素压力34一、耐药性及其发生机制

One.Drugresistanceanditsmechanism

一、耐药性及其发生机制

One.Drugresistan35〔一〕耐药病原体

(One)Drugresistantpathogens革兰氏阴性杆菌：ß-内酰胺酶、超广谱ß-内酰胺酶(ESBL)、Ⅰ类ß-内酰胺酶、多重耐药等；Gram-negativebacillus:ß-lactamase,extendedspectrumß-lactamase(ESBL),Ⅰtypeß-lactamases,multidrugresistance,etc.;革兰氏阳性球菌：耐甲氧西林葡萄球菌(MRS)、耐青霉素肺炎链球菌(PRSP)、耐万古霉素肠球菌(VRE)、高耐氨基糖苷类抗生素肠球菌。Gram-positivecoccus:methicillin-resistantStaphylococcus(MRS),penicillin-resistantStreptococcuspneumoniae(PRSP),vancomycin-resistantenterococcus(VRE),enterococcuswithhighresistancetoaminoglycosideantibiotics.〔一〕耐药病原体

(One)Drugresistant36〔二〕耐药机制

(Two)Drugresistancemechanism1细菌水平和垂直传播耐药基因的整合子系统；1Horizontalandverticaltransmissionofbacterialdrugresistancegenesthroughintegrons;2产生灭活抗生素的水解酶和钝化酶,如ESBLs，AmpCß-内酰胺酶，碳青霉烯酶，氨基糖苷类钝化酶；2proteolyticenzymeandinactiveenzymeproducedforinactivationofantibiotics,suchasESBLs,AmpCß–lactamase,carbapenemase,aminoglycosideinactiveenzyme;〔二〕耐药机制

(Two)Drugresistance373细菌抗生素作用靶位的改变；3Changesinthebacterialantibioticstarget；4细菌膜外排泵出系统；4Bacterialmembraneeffluxpumpsystem；5细菌生物膜的形成。5Bacterialbiofilm(BF)formation.3细菌抗生素作用靶位的改变；38二、检查工程、结果和临床应用examinationprojects,resultsandclinicalapplication二、检查工程、结果和临床应用39〔一〕药物敏感试验

Drugsensitivitytest抗微生物药物敏感试验(antimicrobialsusceptibilitytest,AST)对敏感性不能预测的临床别离菌株进展药敏试验，以指导临床选择治疗药物。ASTcannotbepredictedonthesensitivityofclinicalisolatesforsusceptibilitytestingtoguideclinicaltreatmentofdrug〔一〕药物敏感试验

Drugsensitivitytes40目的

Objective临床别离菌株，如不能对抗生素敏感性进展预测，必须常规进展药敏试验。Clinicalisolates,iftheycannotpredictthesensitivitytoantibioticsshouldberoutinesusceptibilitytesting临床治疗效果差而考虑调整抗菌药物时。Poorclinicaltreatmenttoconsideradjustingtheantimicrobialdrugs.了解细菌耐药的流行病学情况。Understandtheepidemiologyofbacterialresistance.评价新抗菌药物的抗菌谱和抗菌活性等。Evaluationofnewantibioticsandantibacterialactivityofantibacterialspectrum.目的

Objective临床别离菌株，如不能对抗生素敏感性进41方法

Methods1K-B纸片琼脂扩散法〔Kirby-Bauerdiscagardiffusionmethod〕方法

Methods1K-B纸片琼脂扩散法〔Kirby-B42参照NCCLS标准

NCCLSstandardreference敏感〔susceptible，S〕：表示测试菌能被测定药物常规剂量给药后在体内到达的血药浓度所抑制或杀灭。Bacteriacanbedeterminedthatthetestdrugsinthebodyafteradministrationofconventionaldosestoachieveplasmaconcentrationsinhibitorkill.耐药〔resistant，R〕：表示测试菌不能被测定药物常规剂量给药后在体内到达的血药浓度所抑制或杀灭，治疗无效。Testbacteriathatcannotbemeasuredafteradministrationofconventionaldoseofdruginthebodytotheplasmaconcentrationsinhibitorkill,thetreatmentineffective.中介(intermediate，I):该范围作为敏感与耐药之间的缓冲区，防止由于微小技术误差影响实验结果。Therangeasabufferbetweensensitiveandresistanttoavoidtheimpactofthesmalltechnicalerrorresults.参照NCCLS标准

NCCLSstandardrefer432稀释法〔Dilutiontest〕最低抑菌浓度〔MIC〕：能够抑制检测菌肉眼可见生长的最低药物浓度称为测定药物对检测菌的最低抑菌浓度。Minimuminhibitoryconcentration(MIC):Detectionofbacteriaabletoinhibitvisiblegrowthoftheminimumdrugconcentrationdeterminationofdrugsonthedetectionofbacteriaknownastheminimuminhibitoryconcentration.3E试验法〔Etest〕4耐药筛选试验Drugscreeningtest5折点敏感试验Sensitiveturningpointtest2稀释法〔Dilutiontest〕44〔二〕耐药菌监测试验Resistanttomonitortest由于细菌存在一种或几种耐药机制，造成了细菌的多重耐药性。单一的药敏试验已不能完全表示细菌的耐药性，必须进展一些特殊的耐药性监测试验。Becausethereisoneorseveralbacterialresistancemechanisms,resultinginmultipledrugresistanceofbacteria.Singlesusceptibilitytestcannotfullyexpressthedrugresistanceofbacteriatoberesistanttosomespecialmonitoringtests.〔二〕耐药菌监测试验Resistanttomonitor451.耐甲氧西林的葡萄球菌〔methecillinresistancestaphylococcus，MRS〕1μg甲氧西林(苯唑青霉素)纸片的抑菌圈直径≤10mm，或MIC≥4μg/ml的金黄色葡萄球菌,1μg苯唑青霉素纸片的抑菌圈直径≤17mm，或MIC≥0.5μg/ml的凝固酶阴性葡萄球菌称耐甲氧西林葡萄球菌。1μgofmethicillin(oxacillin)paperoftheinhibitionzonediameter≤10mm,orMIC≥4μg/mlStaphylococcusaureus,1μgoxacillindiskinhibitionzonediameter≤17mm,orMIC≥0.5μg/mlofcoagulase-negativemethicillin-resistantStaphylococcusaureus,said1.耐甲氧西林的葡萄球菌〔methecillinresi46临床意义

Clinicalsignificance对所有β-内酰胺类药物均无效,并对氨基糖苷类、大环内酯类、克林霉素和四环素等抗生素多重耐药。治疗首选药物为万古霉素。Forallβ-lactamdrugsareineffective,andaminoglycosides,macrolides,clindamycinandtetracyclineantibioticssuchasmulti-drugresistant.Thepreferreddrugforthetreatmentofvancomycin.临床意义

Clinicalsignificance对所有472.氨基糖苷类抗生素高耐药肠球菌High-resistantenterococciaminoglycosides对氨基糖苷类和青霉素等作用于细胞壁的等抗生素联用无效。Theroleofaminoglycosidesandpenicillinandotherantibioticsinthecellwallcombinedwithnullandvoid.2.氨基糖苷类抗生素高耐药肠球菌High-resistan483.耐青霉素的肺炎链球菌

Penicillin-resistantStreptococcuspneumoniae定义：1μg苯唑青霉素纸片的抑菌圈直径<19mm且MIC>2μg/ml应视为耐青霉素肺炎链球菌〔penicillinresistantstreptococcuspneumonia，PRSP〕。Definition:1μgoxacillindiskinhibitionzonediameter<19mmandMIC>2μg/mlshouldberegardedaspenicillin-resistantStreptococcuspneumoniae3.耐青霉素的肺炎链球菌

Penicillin-resis49临床意义

ClinicalsignificancePRSP对氨苄西林、氨苄西林/舒巴坦、头孢唑啉的临床治疗疗效很差，治疗时参考药敏结果选择药物，经历治疗重症感染时，可用头孢曲松或头孢噻肟联合万古霉素用药。PRSPtoampicillin,ampicillin/sulbactam,cefazolinpoorclinicalefficacyofthetreatmentdrugofchoicewhendrugsusceptibilityresultsforreference,experiencethetreatmentofsevereinfection,canbecombinedceftriaxoneorcefotaximetovancomycintherapy.临床意义

ClinicalsignificancePRSP504.β-内酰胺酶的检测

β-lactamasedetectionβ-内酰胺酶：可水解β-内酰胺类抗生素。β-lactamases:hydrolyzedβ-lactamantibiotics.临床意义：〔1〕流感嗜血杆菌、淋病奈瑟菌以及卡拉莫拉菌等阳性，表示对青霉素、氨苄西林以及阿莫西林耐药。〔2〕葡萄球菌属以及肠球菌属等，阳性表示对青霉素、氨基组青霉素、羧基组青霉素以及脲基组青霉素耐药。Clinicalsignificance:(1),Haemophilusinfluenzae,Neisseriagonorrhoeae,andkaraoke-positivemicroorganismsinMora,saidtopenicillin,ampicillinandamoxicillinresistance.(2)suchasStaphylococcusandEnterococcus,thepositivethatpenicillin,penicillinaminogroup,carboxylgroupofpenicillinandpenicillin-resistanturea-basedgroup.4.β-内酰胺酶的检测

β-lactamasedete515.产超广谱β-内酰胺酶(extendspectrumβ-lactamase,ESBL)的肠杆菌科细菌ProducingESBLEnterobacteriaceae超广谱β-内酰胺酶是由质粒介导的β-内酰胺酶，可水解青霉素类、头孢菌素和氨曲南，主要在大肠埃希菌和克雷伯菌属等肠杆菌科细菌中产生。Extendedspectrumβ-lactamasemediatedbyplasmidβ-lactamases,canbehydrolyzedpenicillins,cephalosporinsandaztreonam,mainlyinEscherichiacoliandKlebsiellaspeciesandotherEnterobacteriaceaeGenerated.5.产超广谱β-内酰胺酶(extendspectrum52临床意义

Clinicalsignificance产ESBL细菌，不管体外药物敏感试验结果如何，对青霉素类、头孢菌素类和氨曲南治疗均无效。ESBLproducingbacteria,regardlessoftheoutcomeofinvitrodrugsensitivitytests,topenicillin,cephalosporinsandaztreonamtreatmentwereineffective.临床意义

Clinicalsignificance产ESB536.耐万古霉素的肠球菌Vancomycinresistantenterococci定义:对30μg万古霉素纸片抑菌圈直径≤19mm，或MIC≥32μg/ml应视为耐万古霉素肠球菌〔vancomycinresistantenterococcus，VRE〕。Definition:paper30μgvancomycininhibitionzonediameter≤19mm,orMIC≥32μg/mlshouldbeconsideredasVRE.6.耐万古霉素的肠球菌Vancomycinresista54临床意义

Clinicalsignificance耐万古霉素肠球菌目前尚无有效的治疗方法，但是对青霉素敏感的VRE可用青霉素和庆大霉素联合治疗，假设对青霉素耐药而不是高水平耐氨基糖苷类可用白霉素和庆大霉素。另外氯霉素、红霉素、四环素〔或多西环素、或米诺环素〕及利福平可用于VRE株。Vancomycinresistantenterococciiscurrentlynoeffectivetreatment,buttheVREtopenicillin-sensitivepenicillinandgentamicincombinationtherapycanbeused,ifnotthehigh-levelresistancetopenicillin-resistantavailableaminoglycosidesneomycinandcelebrategreatwhiteAdriamycin.Additionofchloramphenicol,erythromycinandtetracycline(ordoxycycline,orminocycline)andrifampincanbeusedforVREstrains.临床意义

Clinicalsignificance耐万古霉55〔三〕病原菌耐药基因的检测Detectionofpathogenresistancegenes更早Earlier确证Confirmed准确Accurate金标准Goldstandard〔三〕病原菌耐药基因的检测Detectionofpath56第三节临床感染常见病原体检查Checkthecommonpathogensofclinicalinfection

第三节临床感染常见病原体检查Checkthecommo57感染性疾病指各种生物性病原体〔病原微生物、寄生虫〕寄生人体所引起的传染性感染疾病和非传染性感染疾病。Referstoavarietyofinfectiousdiseases,biologicalpathogens(pathogens,parasites)causedbyhumaninfectiousparasiticdiseasesandnon-communicablediseases.感染性疾病指各种生物性病原体〔病原微生物、寄生虫〕寄生人体所58一、流行病学和临床类型Epidemiologyandclinicaltypes一、流行病学和临床类型Epidemiologyandc59〔一〕流行病学Epidemiology新传染病陆续被发现，老传染病死灰复燃Beendiscoverednewinfectiousdiseases,theresurgenceofolddiseases多重耐药株Multi-drugresistantstrains新医疗技术的开展Thelaunchingofnewmedicaltechnologies〔一〕流行病学Epidemiology新传染病陆续被发现，老60〔二〕临床类型Clinicaltypes临床常见病原体为：细菌、真菌、病毒、支原体、衣原体、螺旋体、立克次体以及寄生虫等。Commonclinicalpathogens:bacteria,fungi,viruses,mycoplasma,chlamydia,spirochetes,rickettsia,andparasites.〔二〕临床类型Clinicaltypes临床常见病原体为：61二、检查工程和临床应用inspectionitemsandclinicalapplications二、检查工程和临床应用inspectionitems62〔一〕细菌感染检查工程的选择

Theselectionofcheckingprojectsonbacterialinfection普通细菌：常采用直接镜检、别离培养与鉴定、抗原检测。Commonbacteria:directmicroscopicexaminationisoftenused,Isolationandidentificationofantigen.病原体抗体检测。Antibodydetectionofpathogens难培养或培养要求高的细菌：综合运用免疫学或分子生物学检测，如结核分枝杆菌感染以及幽门螺杆菌感染等。Difficultforhighcultureorbacterialculture:theintegrateduseofimmunologyormolecularbiology,suchasMycobacteriumtuberculosisandHelicobacterpyloriinfection.细菌培养是最重要确实诊方法。Bacterialcultureisthemostimportantdiagnosticmethod.〔一〕细菌感染检查工程的选择

Theselectiono63〔二〕实验结果分析和临床应用Analysisandclinicalapplicationofexperimentalresults显微镜检查或别离培养的阴性结果不能完全排除感染；Microscopyorisolationandcultureofnegativeresultscannotbecompletelyruledoutinfection共同抗原引起的穿插反响；Antigeniccross-reactivitycausedbythecommon核酸检测的假阳性；False-positivenucleicaciddetection血清学的动态检测。Dynamicdetectionofserological〔二〕实验结果分析和临床应用Analysisandcli64第六节医院感染常见病原体检测Detectionofcommonpathogensofnosocomialinfection

第六节医院感染常见病原体检测Detectionofco65医院感染〔nosocomialinfection〕：又称医院获得性感染〔hospitalacquiredinfection〕，指患者在入院时不存在，也不处于潜伏期，而在医院内发生的感染，包括医院内获得的而在出院后发病的感染。广义的医院感染对象不仅指住院患者，还包括门诊病人、医院职工、探视人员与陪护人员等。Nosocomialinfection(nosocomialinfection):alsoknownashospital-acquiredinfection(hospitalacquiredinfection),thatdoesnotexistonadmissionofpatients,norintheincubationperiod,whileinthehospitalinfectionsoccurred,includingthehospitalafterdischargefromhospitalacquireddiseaseinInfection.Generalizedinfection,notonlybymeansofhospitalinpatients,includingoutpatient,hospitalstaff,visitingofficersandescortpersonnel.医院感染〔nosocomialinfection〕：又称医66一、流行病学和临床类型Epidemiologyandclinicaltypes一、流行病学和临床类型Epidemiologyandc67〔一〕流行病学Epidemiology〔一〕流行病学Epidemiology68病原学

Pathogen细菌是最常见的病原体。Bacterialisthemostcommonpathogens.变化趋势：革兰氏阴性杆菌比例在增加，革兰氏阳性球菌比例在减少。Trend:increasingtheproportionofGram-negativebacteria,Gram-positivecocciratiodecreased.罕见细菌变成流行株：阴沟肠杆菌、洋葱假单孢菌，军团菌等Strainsofbacteriabecomerare:Enterobactercloacae,Pseudomonasonion,Armymonocytogenes其它：病毒、真菌等Other:viruses,fungi,etc病原学

Pathogen细菌是最常见的病原体。692.感染源

Sourceofinfection住院病人、医院职工、探视人员、陪伴人员；Patients,hospitalstaff,visitingstaff,hiscompanion;医院环境、未彻底消毒的医疗器械、血液制品等。Hospitalenvironment,withoutthoroughdisinfectionofmedicaldevices,bloodproducts,etc.2.感染源

Sourceofinfection住院病人70高危人群

High-riskgroups慢性疾病患者，如肿瘤、糖尿病以及肝硬化患者等Patientswithchronicdiseasessuchascancer,diabetesandlivercirrhosis,etc.

免疫抑制剂及激素的使用者Immunosuppressivedrugsandhormoneusers

化疗及放疗患者Chemotherapyandradiotherapyinpatients

手术及麻醉患者Surgeryandanesthesiainpatients

使用介入性诊断与治疗的患者Useofinterventionaldiagnosisandtreatmentofpatients

广谱抗生素使用者Broad-spectrumantibioticusers高危人群

High-riskgroups慢性疾病患者，如肿71〔二〕常见临床类型

Commonclinicaltypes1下呼吸道感染：最常见Lowerrespiratorytractinfection:themostcommon2尿路感染UrinaryTractInfection3手术切口感染：外源性和内源性Surgicalwoundinfection:exogenousandendogenous4胃肠道感染Gastrointestinalinfections5血液感染Bloodinfection6皮肤和软组织感染Skinandsofttissueinfections〔二〕常见临床类型

Commonclinicaltype72二、检查工程和临床应用inspectionitemsandclinicalapplications二、检查工程和临床应用inspectionitemsa73〔一〕医院感染病原体检查工程和临床应用inspectionitemsandclinicalapplicationsforHospitalInfection〔一〕医院感染病原体检查工程和临床应用74根本原那么与病原体感染类似，但更应注意正常菌群的污染以及条件致病菌在医院感染中的地位。Similartothebasicprinciplesandpathogeninfection,butmoreshouldpayattentiontothepollutionofthenormalfloraandopportunisticpathogensinthehospitalinfectionstatus.根本原那么与病原体感染类似，但更应注意正常菌群的污染以及条件75标本采集和送检根本原那么

Thebasicprinciplesofspecimencollectionandsubmission及时采集标本Collectspecimensinatimelymanner严格无菌操作Strictaseptic立即送出Immediatelysent在抗菌药物使用前Beforetheuseofantibiotics适宜的容器Appropriatecontainer注明来源和目的Indicatethesourceanddestination定量或半定量培养Quantitativeorsemi-quantitativeculture分级报告和限时报告Reportsandlimitgradingreport标本采集和送检根本原那么

Thebasicprincip762.涂片镜检

Smearmicroscopy痰标本Sputum2.涂片镜检

Smearmicroscopy痰标本773.别离培养鉴定法

Isolationandcultureidentificationmethod尿路感染UrinaryTractInfection手术切口感染Surgicalwoundinfection大便培养Stoolculture血培养Bloodculture3.别离培养鉴定法

Isolationandcultu78〔二〕医院环境中细菌污染的监测和消毒灭菌效果的监测Bacterialcontaminationinthehospitalenvironmentmonitoringandthemonitoringofsterilizationresults〔二〕医院环境中细菌污染的监测和消毒灭菌效果的监测79定期对医院环境进展病原体监测：空气、物体外表、医务人员手部；Regularmonitoringofpathogensinhospitalenvironment:air,surface,themedicalstaffofhand;定期对医疗器械的消毒效果进展监测：高压蒸气、紫外线、化学消毒剂。Regulardisinfectionofmedicaldevicestomonitor:high-pressuresteam,UVlight,chemicaldisinfectants.定期对医院环境进展病原体监测：空气、物体外表、医务人员手部；80Thanks！Thanks！81临床常见病原体检测

Examinationofclinicalcommonpathogens呼吸内科熊维宁DepartmentofRespiratoryMedicineXiong,Weining临床常见病原体检测

Examinationofclin82目的

Objective确定感染的发生和性质，及早明确诊断；Determinetheoccuringandnatureofinfection,andmakediagnosisearly;尽早选择适当的治疗方案；Selecttheappropriatetreatmentassoonaspossible;采取有效的预防措施，防止感染可能广泛传播所造成的危害。Takeeffectivepreventionmeasurestopreventinfectiontocausewidespreaddamage.目的

Objective确定感染的发生和性质，及早明确诊断；83根本程序

BasicProcedures正确采集和运送标本；Correctcollectionandtransportationofspecimens初步诊断：直接镜检，免疫学和分子生物学检测；Initialdiagnosis:directmicroscopy,immunologyandmolecularbiologyexamination;确定诊断：病原体的别离、鉴定及药敏实验，报告结果；Determineddiagnosis:pathogenisolation,identificationandsensitivitytest,reportingresults;合理用药Appropriatedruguse根本程序

BasicProcedures正确采集和运送标本84第一节

SectionOne标本的采集运送、实验室评价和检查方法Transportationandcollectionofspecimen,laboratoryevaluationandexaminationmethods第一节

SectionOne标本的采集运送、实验室评价和检85一、标本采集和运送One.CollectionandtransportationofSpecimen一、标本采集和运送86根本原那么

Basicprinciples根据病史与临床表现确定标本采集的时间、部位、种类和数量，尽量采集病变明显部位标本；Accordingtohistoryandclinicalmanifestationstodeterminethecollectiontime,location,typeandquantityofspecimens,collectspecimensatsignificantpartsoflesions;无菌操作，防止污染；Useaseptictechniquetopreventcontamination;采取适宜的方式进展储存，尽快送检；Appropriatewaytostore,assoonaspossibleforexamination;要视所有标本为传染品，高度危险性的标本要有明显标识，急症或危重患者标本要特别注明。Toviewallthespecimensforinfectious,highriskspecimensshouldbeclearlymarked,acuteorcriticallyillpatientspecimensshouldbespecified.根本原那么

Basicprinciples根据病史与临床表87临床常见病原体检测英文版教学课件88临床常见病原体检测英文版教学课件89〔二〕尿液

(Two)Urine无菌采集中段尿；Sterilecollectionofmid-portionurine;如考虑厌氧菌感染，采取膀胱穿刺法采集标本，无菌厌氧小瓶运送；Ifconsiderationofanaerobicinfections,applybladderpuncturetocollectspecimens,transportthemwithsterileanaerobicvial;排尿困难者考虑导尿采集标本。Applycatheterizationtocollectspecimensfordysuriapatients.〔二〕尿液

(Two)Urine无菌采集中段尿；90〔三〕粪便

(Three)Stool挑取脓、血或粘液局部于清洁容器中送检；Pickpus,bloodormucusfromstollintocleancontainersforexamination;排便困难者或婴儿采用直肠拭子采集，置于有保存液的试管内送检；Applyrectalswabforinfantsordifficultdefecation,placeitwithpreservationsolutioninthetesttubeforexamination;疑心霍乱弧菌感染引起的腹泻，将标本置于碱性蛋白胨水或卡-布〔Cary-Blair〕运送培养液送检；SuspectedinfectionscausedbyVibriocholerae,thespecimenswereplacedinalkalinepeptonewaterorcard-cloth(Cary-Blair)transportmediumforexamination;传染性腹泻应连续送检3次。Continuousexamination3timesforinfectiousdiarrhea.〔三〕粪便

(Three)Stool挑取脓、血或粘液局部于91〔四〕呼吸道标本

(Four)Respiratoryspecimens类型：鼻咽拭子，痰和经气管采集的标本；Types:nasopharyngealswab,sputum,andspecimenscollectedthroughtracheal上呼吸道存在正常菌群，在采集标本与结果分析时应予考虑。Thereisnormalflorainupperrespiratorytract,thespecimenscollectionandresultsanalysisshouldbeconsidered.〔四〕呼吸道标本

(Four)Respiratorysp92〔五〕脑脊液与其他无菌体液

(Five)Cerebrospinalfluidandothersterilebodyfluids脑脊液应立即保温送检或床边接种；CSFshouldbetransportedimmediatelywithinsulationorculturedbedside；胸腔积液、腹腔积液和心包积液等应采集较大量标本送检，离心后再接种培养。Pleuraleffusion,ascitesandpericardialeffusionwithalargevolumeshouldbecollected,centrifugatedandthencultured.〔五〕脑脊液与其他无菌体液

(Five)Cerebrosp93〔六〕眼、耳部标本

(Six)Eye,earspecimens拭子采样Swabsamples〔六〕眼、耳部标本

(Six)Eye,earspeci94〔七〕泌尿生殖道标本

(Seven)Urogenitalspecimens男性：无菌采集尿道口分泌物或前列腺液；Male:urethraorprostaticfluidsecretionswithsterilecollection;女性：无菌采集阴道或宫颈分泌物。Female:vaginalorcervicalsecretionscollectedaseptically.〔七〕泌尿生殖道标本

(Seven)Urogenital95〔八〕创伤、组织和脓肿标本

(Eight)Trauma,organization,andabscessspecimens对损伤范围较大的创伤，从不同部位采集多份标本；Onawiderrangeoftraumainjuries,specimenscollectedfrommanydifferentparts;采集部位应首先去除污物、消毒皮肤；Firstlyremovedirt,disinfecttheskinaroundcollectionsite;标本较少那么需参加无菌生理盐水以防枯燥；Fewersamplesmustbeaddedtosterilesalinetopreventdrying;开放性脓肿及脓性分泌物：用无菌棉纤采取脓液或病灶深局部泌物；Openabscessandpurulentsecretions:collectdeepdischargepusorlesionswithsterilecottonfibers;封闭性脓肿用注射器抽取；Collectspecimeninclosedabscesswithasyringe;疑心厌氧菌感染，应隔绝空气采集。Suspectedanaerobicinfections,collectionshouldbeisolatedfromair.〔八〕创伤、组织和脓肿标本

(Eight)Trauma,96〔九〕血清

(Nine)Serum用于检测特异性抗体；Forthedetectionofspecificantibodies;血液自然凝固后吸取血清，灭活补体。Drawserumafternaturalcoagulationofblood,theninactivatecomplement.〔九〕血清

(Nine)Serum用于检测特异性抗体；97二、标本的实验室质量评估标准

Two.Standardoflaboratoryqualityassessmentofspecimens二、标本的实验室质量评估标准

Two.Standardo98检验申请单的根本内容

ThebasiccontentofanapplicationforexaminationT