臨床常見病原體檢測(cè)英文版教學(xué)課件

臨床常見病原體檢測(cè)

Examinationofclinicalcommonpathogens呼吸內(nèi)科熊維寧DepartmentofRespiratoryMedicineXiong,Weining臨床常見病原體檢測(cè)

Examinationofclin1目的

Objective確定感染的發(fā)生和性質(zhì)，及早明確診斷；Determinetheoccuringandnatureofinfection,andmakediagnosisearly;盡早選擇適當(dāng)?shù)闹委煼桨?；Selecttheappropriatetreatmentassoonaspossible;采取有效的預(yù)防措施，防止感染可能廣泛傳播所造成的危害。Takeeffectivepreventionmeasurestopreventinfectiontocausewidespreaddamage.目的

Objective確定感染的發(fā)生和性質(zhì)，及早明確診斷；2根本程序

BasicProcedures正確采集和運(yùn)送標(biāo)本；Correctcollectionandtransportationofspecimens初步診斷：直接鏡檢，免疫學(xué)和分子生物學(xué)檢測(cè)；Initialdiagnosis:directmicroscopy,immunologyandmolecularbiologyexamination;確定診斷：病原體的別離、鑒定及藥敏實(shí)驗(yàn)，報(bào)告結(jié)果；Determineddiagnosis:pathogenisolation,identificationandsensitivitytest,reportingresults;合理用藥Appropriatedruguse根本程序

BasicProcedures正確采集和運(yùn)送標(biāo)本3第一節(jié)

SectionOne標(biāo)本的采集運(yùn)送、實(shí)驗(yàn)室評(píng)價(jià)和檢查方法Transportationandcollectionofspecimen,laboratoryevaluationandexaminationmethods第一節(jié)

SectionOne標(biāo)本的采集運(yùn)送、實(shí)驗(yàn)室評(píng)價(jià)和檢4一、標(biāo)本采集和運(yùn)送One.CollectionandtransportationofSpecimen一、標(biāo)本采集和運(yùn)送5根本原那么

Basicprinciples根據(jù)病史與臨床表現(xiàn)確定標(biāo)本采集的時(shí)間、部位、種類和數(shù)量，盡量采集病變明顯部位標(biāo)本；Accordingtohistoryandclinicalmanifestationstodeterminethecollectiontime,location,typeandquantityofspecimens,collectspecimensatsignificantpartsoflesions;無菌操作，防止污染；Useaseptictechniquetopreventcontamination;采取適宜的方式進(jìn)展儲(chǔ)存，盡快送檢；Appropriatewaytostore,assoonaspossibleforexamination;要視所有標(biāo)本為傳染品，高度危險(xiǎn)性的標(biāo)本要有明顯標(biāo)識(shí)，急癥或危重患者標(biāo)本要特別注明。Toviewallthespecimensforinfectious,highriskspecimensshouldbeclearlymarked,acuteorcriticallyillpatientspecimensshouldbespecified.根本原那么

Basicprinciples根據(jù)病史與臨床表6臨床常見病原體檢測(cè)英文版教學(xué)課件7臨床常見病原體檢測(cè)英文版教學(xué)課件8〔二〕尿液

(Two)Urine無菌采集中段尿；Sterilecollectionofmid-portionurine;如考慮厭氧菌感染，采取膀胱穿刺法采集標(biāo)本，無菌厭氧小瓶運(yùn)送；Ifconsiderationofanaerobicinfections,applybladderpuncturetocollectspecimens,transportthemwithsterileanaerobicvial;排尿困難者考慮導(dǎo)尿采集標(biāo)本。Applycatheterizationtocollectspecimensfordysuriapatients.〔二〕尿液

(Two)Urine無菌采集中段尿；9〔三〕糞便

(Three)Stool挑取膿、血或粘液局部于清潔容器中送檢；Pickpus,bloodormucusfromstollintocleancontainersforexamination;排便困難者或嬰兒采用直腸拭子采集，置于有保存液的試管內(nèi)送檢；Applyrectalswabforinfantsordifficultdefecation,placeitwithpreservationsolutioninthetesttubeforexamination;疑心霍亂弧菌感染引起的腹瀉，將標(biāo)本置于堿性蛋白胨水或卡-布〔Cary-Blair〕運(yùn)送培養(yǎng)液送檢；SuspectedinfectionscausedbyVibriocholerae,thespecimenswereplacedinalkalinepeptonewaterorcard-cloth(Cary-Blair)transportmediumforexamination;傳染性腹瀉應(yīng)連續(xù)送檢3次。Continuousexamination3timesforinfectiousdiarrhea.〔三〕糞便

(Three)Stool挑取膿、血或粘液局部于10〔四〕呼吸道標(biāo)本

(Four)Respiratoryspecimens類型：鼻咽拭子，痰和經(jīng)氣管采集的標(biāo)本；Types:nasopharyngealswab,sputum,andspecimenscollectedthroughtracheal上呼吸道存在正常菌群，在采集標(biāo)本與結(jié)果分析時(shí)應(yīng)予考慮。Thereisnormalflorainupperrespiratorytract,thespecimenscollectionandresultsanalysisshouldbeconsidered.〔四〕呼吸道標(biāo)本

(Four)Respiratorysp11〔五〕腦脊液與其他無菌體液

(Five)Cerebrospinalfluidandothersterilebodyfluids腦脊液應(yīng)立即保溫送檢或床邊接種；CSFshouldbetransportedimmediatelywithinsulationorculturedbedside；胸腔積液、腹腔積液和心包積液等應(yīng)采集較大量標(biāo)本送檢，離心后再接種培養(yǎng)。Pleuraleffusion,ascitesandpericardialeffusionwithalargevolumeshouldbecollected,centrifugatedandthencultured.〔五〕腦脊液與其他無菌體液

(Five)Cerebrosp12〔六〕眼、耳部標(biāo)本

(Six)Eye,earspecimens拭子采樣Swabsamples〔六〕眼、耳部標(biāo)本

(Six)Eye,earspeci13〔七〕泌尿生殖道標(biāo)本

(Seven)Urogenitalspecimens男性：無菌采集尿道口分泌物或前列腺液；Male:urethraorprostaticfluidsecretionswithsterilecollection;女性：無菌采集陰道或?qū)m頸分泌物。Female:vaginalorcervicalsecretionscollectedaseptically.〔七〕泌尿生殖道標(biāo)本

(Seven)Urogenital14〔八〕創(chuàng)傷、組織和膿腫標(biāo)本

(Eight)Trauma,organization,andabscessspecimens對(duì)損傷范圍較大的創(chuàng)傷，從不同部位采集多份標(biāo)本；Onawiderrangeoftraumainjuries,specimenscollectedfrommanydifferentparts;采集部位應(yīng)首先去除污物、消毒皮膚；Firstlyremovedirt,disinfecttheskinaroundcollectionsite;標(biāo)本較少那么需參加無菌生理鹽水以防枯燥；Fewersamplesmustbeaddedtosterilesalinetopreventdrying;開放性膿腫及膿性分泌物：用無菌棉纖采取膿液或病灶深局部泌物；Openabscessandpurulentsecretions:collectdeepdischargepusorlesionswithsterilecottonfibers;封閉性膿腫用注射器抽?。籆ollectspecimeninclosedabscesswithasyringe;疑心厭氧菌感染，應(yīng)隔絕空氣采集。Suspectedanaerobicinfections,collectionshouldbeisolatedfromair.〔八〕創(chuàng)傷、組織和膿腫標(biāo)本

(Eight)Trauma,15〔九〕血清

(Nine)Serum用于檢測(cè)特異性抗體；Forthedetectionofspecificantibodies;血液自然凝固后吸取血清，滅活補(bǔ)體。Drawserumafternaturalcoagulationofblood,theninactivatecomplement.〔九〕血清

(Nine)Serum用于檢測(cè)特異性抗體；16二、標(biāo)本的實(shí)驗(yàn)室質(zhì)量評(píng)估標(biāo)準(zhǔn)

Two.Standardoflaboratoryqualityassessmentofspecimens二、標(biāo)本的實(shí)驗(yàn)室質(zhì)量評(píng)估標(biāo)準(zhǔn)

Two.Standardo17檢驗(yàn)申請(qǐng)單的根本內(nèi)容

ThebasiccontentofanapplicationforexaminationThepatient,snameandhospitalnumber.Ageandsex.Collectiondateandtime.Suspecteddiagnosis.Exactnatureandsourceofthespecimen.Immunizationhistoryandantimicrobialtherapy.Objective.Signedbyphysician.檢驗(yàn)申請(qǐng)單的根本內(nèi)容

Thebasiccontento18標(biāo)本接收和拒收準(zhǔn)那么

Receptionandrejectioncriteriaforspecimens1Theinformationonthelabeldoesnotexistornotmatchtheinformationontherequisition;2Delayindeliveryofthespecimens;3Thespecimenisleaking;4Thespecimenhasbeentransportedattheimpropertemperatureorinimpropermedium.標(biāo)本接收和拒收準(zhǔn)那么

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

Three.Examinationmethods

三、檢查方法

Three.Examinationmeth21〔一〕直接顯微鏡檢查

(One)Directmicroscopicexamination不染色標(biāo)本檢查法：用于觀察病原體的生長(zhǎng)、形態(tài)與運(yùn)動(dòng)等特性；Notstainedspecimentestmethod:toobservethegrowth,shapeandmotioncharacteristicsofpathogens;評(píng)價(jià)：局部病原體可借此初步診斷。Evaluation:forsomepathogens,theinitialdiagnosiscanbetaken.〔一〕直接顯微鏡檢查

(One)Directmicros22染色標(biāo)本檢查法：觀察細(xì)菌的形態(tài)、染色性或觀察宿主細(xì)胞內(nèi)包涵體的特征；Stainedspecimentestmethod:toobservebacterialmorphology,stainingorobservethecharacteristicsoftheinclusionbodiesinhostcell;評(píng)價(jià)：為臨床初步診斷提供依據(jù)。Evaluation:availablebasisforthepreliminaryclinicaldiagnosis.染色標(biāo)本檢查法：觀察細(xì)菌的形態(tài)、染色性或觀察宿主細(xì)胞內(nèi)包涵體23革蘭氏〔染色〕陽(yáng)性球菌Gram(staining)-positivecocci革蘭氏〔染色〕陰性桿菌Gram(staining)-negativebacilli革蘭氏〔染色〕陽(yáng)性球菌革蘭氏〔染色〕陰性桿菌24抗酸染色陽(yáng)性桿菌〔抗酸桿菌〕Acid-faststain-positivebacilli(Acid-fastbacilli)抗酸染色陽(yáng)性桿菌〔抗酸桿菌〕25〔二〕病原體特異性抗原檢查

(Two)Pathogen-specificantigentest免疫熒光技術(shù)Immunofluorescencetechnique酶聯(lián)免疫技術(shù)ELISA化學(xué)發(fā)光技術(shù)Chemiluminescencetechnology乳膠凝集試驗(yàn)Latexagglutinationtest對(duì)流免疫電泳Counterimmunoelectrophoresis蛋白質(zhì)芯片Proteinchips〔二〕病原體特異性抗原檢查

(Two)Pathogen-s26病原體特異性抗原檢測(cè)結(jié)果評(píng)價(jià)

Evaluationofpathogen-specificantigentestresults如果能排除穿插抗原的影響，病原體抗原檢測(cè)可明確感染的病原體。Iftheimpactofcross-antigencanbeexcluded,pathogenantigentestcouldcleartheinfectionofpathogens.病原體特異性抗原檢測(cè)結(jié)果評(píng)價(jià)

Evaluationofp27〔三〕病原體核酸檢查

(Three)Pathogennucleicacidtest聚合酶鏈?zhǔn)椒错懀簲U(kuò)增病原體微生物特異的DNA或RNA片段；Polymerasechainreaction(PCR):amplifythespecificfragmentsofDNAorRNAofmicrobialpathogens;實(shí)時(shí)熒光定量PCR技術(shù)；Real-timePCR;核酸探針雜交技術(shù)：通過序列的探針與標(biāo)本中的病原體的核苷酸雜交，用以了解病原體的有無；Nucleicacidprobehybridization:hybridizationbetweentheknownsequenceoftheprobeandthenucleotideofpathogensinthesamples,toexplorewhetherpathogensexist;基因芯片技術(shù)。GenechiporDNAmicroarray.〔三〕病原體核酸檢查

(Three)Pathogennu28病原體核酸檢查結(jié)果評(píng)價(jià)

EvaluationofpathogenDNAtestresults是檢測(cè)病原體微生物最靈敏的方法，但具有一定的假陽(yáng)性與假陰性；isthemostsensitivedetectionmethodformicrobialpathogens,buthassomefalsepositiveandfalsenegative;陽(yáng)性只說明存在某種病原體的核酸，是否正被感染應(yīng)結(jié)合臨床具體分析。istheonlyshowthattheexistenceofapositivepathogennucleicacid,whetherbeinginfectedshouldbecombinedwithclinicalspecificanalysis.病原體核酸檢查結(jié)果評(píng)價(jià)

Evaluationofpath29〔四〕病原體的別離培養(yǎng)與鑒定

(Four)Isolation,cultureandidentificationofpathogens1細(xì)菌感染性疾病病原體的別離培養(yǎng)1Isolationandculturethepathogensofbacterialinfections明確感染病原體；Clearthepathogens;為臨床提供體外抗微生物藥物敏感試驗(yàn)結(jié)果。Providevitrotestresultsofanti-microbialdrugsensitivityforclinical.〔四〕病原體的別離培養(yǎng)與鑒定

(Four)Isolatio302不能人工培養(yǎng)的病原體感染性疾病2Notcultivatedpathogensofinfectiousdisease將標(biāo)本接種易感動(dòng)物、雞胚或行細(xì)胞培養(yǎng)。Specimenswereinoculatedintosusceptibleanimals,eggsorcelllines.2不能人工培養(yǎng)的病原體感染性疾病31〔五〕血清學(xué)實(shí)驗(yàn)

(Five)Serologicaltest特異性IgM可作為感染性疾病的早期診斷指標(biāo)，且可區(qū)分原發(fā)與復(fù)發(fā)感染；SpecificIgMcanbeindicatorofearlydiagnosisforinfectiousdiseasesandcandistinguishbetweenprimaryandrecurrentinfections;特異性IgG，尤其雙份血清的滴度呈4倍或4倍以上升高，考慮現(xiàn)癥感染。ForIgG,thetiterofdoubleserumhigher4timesormorethan4times,currentpathogeninfectionshouldbeconsidered.〔五〕血清學(xué)實(shí)驗(yàn)

(Five)Serologicalte32第二節(jié)病原體耐藥性檢測(cè)

SectiontwoDetectionofpathogendrugresistance

第二節(jié)病原體耐藥性檢測(cè)

SectiontwoDetec33抗生素壓力Antibioticpressure抗生素壓力34一、耐藥性及其發(fā)生機(jī)制

One.Drugresistanceanditsmechanism

一、耐藥性及其發(fā)生機(jī)制

One.Drugresistan35〔一〕耐藥病原體

(One)Drugresistantpathogens革蘭氏陰性桿菌：?-內(nèi)酰胺酶、超廣譜?-內(nèi)酰胺酶(ESBL)、Ⅰ類?-內(nèi)酰胺酶、多重耐藥等；Gram-negativebacillus:?-lactamase,extendedspectrum?-lactamase(ESBL),Ⅰtype?-lactamases,multidrugresistance,etc.;革蘭氏陽(yáng)性球菌：耐甲氧西林葡萄球菌(MRS)、耐青霉素肺炎鏈球菌(PRSP)、耐萬古霉素腸球菌(VRE)、高耐氨基糖苷類抗生素腸球菌。Gram-positivecoccus:methicillin-resistantStaphylococcus(MRS),penicillin-resistantStreptococcuspneumoniae(PRSP),vancomycin-resistantenterococcus(VRE),enterococcuswithhighresistancetoaminoglycosideantibiotics.〔一〕耐藥病原體

(One)Drugresistant36〔二〕耐藥機(jī)制

(Two)Drugresistancemechanism1細(xì)菌水平和垂直傳播耐藥基因的整合子系統(tǒng)；1Horizontalandverticaltransmissionofbacterialdrugresistancegenesthroughintegrons;2產(chǎn)生滅活抗生素的水解酶和鈍化酶,如ESBLs，AmpC?-內(nèi)酰胺酶，碳青霉烯酶，氨基糖苷類鈍化酶；2proteolyticenzymeandinactiveenzymeproducedforinactivationofantibiotics,suchasESBLs,AmpC?–lactamase,carbapenemase,aminoglycosideinactiveenzyme;〔二〕耐藥機(jī)制

(Two)Drugresistance373細(xì)菌抗生素作用靶位的改變；3Changesinthebacterialantibioticstarget；4細(xì)菌膜外排泵出系統(tǒng)；4Bacterialmembraneeffluxpumpsystem；5細(xì)菌生物膜的形成。5Bacterialbiofilm(BF)formation.3細(xì)菌抗生素作用靶位的改變；38二、檢查工程、結(jié)果和臨床應(yīng)用examinationprojects,resultsandclinicalapplication二、檢查工程、結(jié)果和臨床應(yīng)用39〔一〕藥物敏感試驗(yàn)

Drugsensitivitytest抗微生物藥物敏感試驗(yàn)(antimicrobialsusceptibilitytest,AST)對(duì)敏感性不能預(yù)測(cè)的臨床別離菌株進(jìn)展藥敏試驗(yàn)，以指導(dǎo)臨床選擇治療藥物。ASTcannotbepredictedonthesensitivityofclinicalisolatesforsusceptibilitytestingtoguideclinicaltreatmentofdrug〔一〕藥物敏感試驗(yàn)

Drugsensitivitytes40目的

Objective臨床別離菌株，如不能對(duì)抗生素敏感性進(jìn)展預(yù)測(cè)，必須常規(guī)進(jìn)展藥敏試驗(yàn)。Clinicalisolates,iftheycannotpredictthesensitivitytoantibioticsshouldberoutinesusceptibilitytesting臨床治療效果差而考慮調(diào)整抗菌藥物時(shí)。Poorclinicaltreatmenttoconsideradjustingtheantimicrobialdrugs.了解細(xì)菌耐藥的流行病學(xué)情況。Understandtheepidemiologyofbacterialresistance.評(píng)價(jià)新抗菌藥物的抗菌譜和抗菌活性等。Evaluationofnewantibioticsandantibacterialactivityofantibacterialspectrum.目的

Objective臨床別離菌株，如不能對(duì)抗生素敏感性進(jìn)41方法

Methods1K-B紙片瓊脂擴(kuò)散法〔Kirby-Bauerdiscagardiffusionmethod〕方法

Methods1K-B紙片瓊脂擴(kuò)散法〔Kirby-B42參照NCCLS標(biāo)準(zhǔn)

NCCLSstandardreference敏感〔susceptible，S〕：表示測(cè)試菌能被測(cè)定藥物常規(guī)劑量給藥后在體內(nèi)到達(dá)的血藥濃度所抑制或殺滅。Bacteriacanbedeterminedthatthetestdrugsinthebodyafteradministrationofconventionaldosestoachieveplasmaconcentrationsinhibitorkill.耐藥〔resistant，R〕：表示測(cè)試菌不能被測(cè)定藥物常規(guī)劑量給藥后在體內(nèi)到達(dá)的血藥濃度所抑制或殺滅，治療無效。Testbacteriathatcannotbemeasuredafteradministrationofconventionaldoseofdruginthebodytotheplasmaconcentrationsinhibitorkill,thetreatmentineffective.中介(intermediate，I):該范圍作為敏感與耐藥之間的緩沖區(qū)，防止由于微小技術(shù)誤差影響實(shí)驗(yàn)結(jié)果。Therangeasabufferbetweensensitiveandresistanttoavoidtheimpactofthesmalltechnicalerrorresults.參照NCCLS標(biāo)準(zhǔn)

NCCLSstandardrefer432稀釋法〔Dilutiontest〕最低抑菌濃度〔MIC〕：能夠抑制檢測(cè)菌肉眼可見生長(zhǎng)的最低藥物濃度稱為測(cè)定藥物對(duì)檢測(cè)菌的最低抑菌濃度。Minimuminhibitoryconcentration(MIC):Detectionofbacteriaabletoinhibitvisiblegrowthoftheminimumdrugconcentrationdeterminationofdrugsonthedetectionofbacteriaknownastheminimuminhibitoryconcentration.3E試驗(yàn)法〔Etest〕4耐藥篩選試驗(yàn)Drugscreeningtest5折點(diǎn)敏感試驗(yàn)Sensitiveturningpointtest2稀釋法〔Dilutiontest〕44〔二〕耐藥菌監(jiān)測(cè)試驗(yàn)Resistanttomonitortest由于細(xì)菌存在一種或幾種耐藥機(jī)制，造成了細(xì)菌的多重耐藥性。單一的藥敏試驗(yàn)已不能完全表示細(xì)菌的耐藥性，必須進(jìn)展一些特殊的耐藥性監(jiān)測(cè)試驗(yàn)。Becausethereisoneorseveralbacterialresistancemechanisms,resultinginmultipledrugresistanceofbacteria.Singlesusceptibilitytestcannotfullyexpressthedrugresistanceofbacteriatoberesistanttosomespecialmonitoringtests.〔二〕耐藥菌監(jiān)測(cè)試驗(yàn)Resistanttomonitor451.耐甲氧西林的葡萄球菌〔methecillinresistancestaphylococcus，MRS〕1μg甲氧西林(苯唑青霉素)紙片的抑菌圈直徑≤10mm，或MIC≥4μg/ml的金黃色葡萄球菌,1μg苯唑青霉素紙片的抑菌圈直徑≤17mm，或MIC≥0.5μg/ml的凝固酶陰性葡萄球菌稱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臨床意義

Clinicalsignificance對(duì)所有β-內(nèi)酰胺類藥物均無效,并對(duì)氨基糖苷類、大環(huán)內(nèi)酯類、克林霉素和四環(huán)素等抗生素多重耐藥。治療首選藥物為萬古霉素。Forallβ-lactamdrugsareineffective,andaminoglycosides,macrolides,clindamycinandtetracyclineantibioticssuchasmulti-drugresistant.Thepreferreddrugforthetreatmentofvancomycin.臨床意義

Clinicalsignificance對(duì)所有472.氨基糖苷類抗生素高耐藥腸球菌High-resistantenterococciaminoglycosides對(duì)氨基糖苷類和青霉素等作用于細(xì)胞壁的等抗生素聯(lián)用無效。Theroleofaminoglycosidesandpenicillinandotherantibioticsinthecellwallcombinedwithnullandvoid.2.氨基糖苷類抗生素高耐藥腸球菌High-resistan483.耐青霉素的肺炎鏈球菌

Penicillin-resistantStreptococcuspneumoniae定義：1μg苯唑青霉素紙片的抑菌圈直徑<19mm且MIC>2μg/ml應(yīng)視為耐青霉素肺炎鏈球菌〔penicillinresistantstreptococcuspneumonia，PRSP〕。Definition:1μgoxacillindiskinhibitionzonediameter<19mmandMIC>2μg/mlshouldberegardedaspenicillin-resistantStreptococcuspneumoniae3.耐青霉素的肺炎鏈球菌

Penicillin-resis49臨床意義

ClinicalsignificancePRSP對(duì)氨芐西林、氨芐西林/舒巴坦、頭孢唑啉的臨床治療療效很差，治療時(shí)參考藥敏結(jié)果選擇藥物，經(jīng)歷治療重癥感染時(shí)，可用頭孢曲松或頭孢噻肟聯(lián)合萬古霉素用藥。PRSPtoampicillin,ampicillin/sulbactam,cefazolinpoorclinicalefficacyofthetreatmentdrugofchoicewhendrugsusceptibilityresultsforreference,experiencethetreatmentofsevereinfection,canbecombinedceftriaxoneorcefotaximetovancomycintherapy.臨床意義

ClinicalsignificancePRSP504.β-內(nèi)酰胺酶的檢測(cè)

β-lactamasedetectionβ-內(nèi)酰胺酶：可水解β-內(nèi)酰胺類抗生素。β-lactamases:hydrolyzedβ-lactamantibiotics.臨床意義：〔1〕流感嗜血桿菌、淋病奈瑟菌以及卡拉莫拉菌等陽(yáng)性，表示對(duì)青霉素、氨芐西林以及阿莫西林耐藥。〔2〕葡萄球菌屬以及腸球菌屬等，陽(yáng)性表示對(duì)青霉素、氨基組青霉素、羧基組青霉素以及脲基組青霉素耐藥。Clinicalsignificance:(1),Haemophilusinfluenzae,Neisseriagonorrhoeae,andkaraoke-positivemicroorganismsinMora,saidtopenicillin,ampicillinandamoxicillinresistance.(2)suchasStaphylococcusandEnterococcus,thepositivethatpenicillin,penicillinaminogroup,carboxylgroupofpenicillinandpenicillin-resistanturea-basedgroup.4.β-內(nèi)酰胺酶的檢測(cè)

β-lactamasedete515.產(chǎn)超廣譜β-內(nèi)酰胺酶(extendspectrumβ-lactamase,ESBL)的腸桿菌科細(xì)菌ProducingESBLEnterobacteriaceae超廣譜β-內(nèi)酰胺酶是由質(zhì)粒介導(dǎo)的β-內(nèi)酰胺酶，可水解青霉素類、頭孢菌素和氨曲南，主要在大腸埃希菌和克雷伯菌屬等腸桿菌科細(xì)菌中產(chǎn)生。Extendedspectrumβ-lactamasemediatedbyplasmidβ-lactamases,canbehydrolyzedpenicillins,cephalosporinsandaztreonam,mainlyinEscherichiacoliandKlebsiellaspeciesandotherEnterobacteriaceaeGenerated.5.產(chǎn)超廣譜β-內(nèi)酰胺酶(extendspectrum52臨床意義

Clinicalsignificance產(chǎn)ESBL細(xì)菌，不管體外藥物敏感試驗(yàn)結(jié)果如何，對(duì)青霉素類、頭孢菌素類和氨曲南治療均無效。ESBLproducingbacteria,regardlessoftheoutcomeofinvitrodrugsensitivitytests,topenicillin,cephalosporinsandaztreonamtreatmentwereineffective.臨床意義

Clinicalsignificance產(chǎn)ESB536.耐萬古霉素的腸球菌Vancomycinresistantenterococci定義:對(duì)30μg萬古霉素紙片抑菌圈直徑≤19mm，或MIC≥32μg/ml應(yīng)視為耐萬古霉素腸球菌〔vancomycinresistantenterococcus，VRE〕。Definition:paper30μgvancomycininhibitionzonediameter≤19mm,orMIC≥32μg/mlshouldbeconsideredasVRE.6.耐萬古霉素的腸球菌Vancomycinresista54臨床意義

Clinicalsignificance耐萬古霉素腸球菌目前尚無有效的治療方法，但是對(duì)青霉素敏感的VRE可用青霉素和慶大霉素聯(lián)合治療，假設(shè)對(duì)青霉素耐藥而不是高水平耐氨基糖苷類可用白霉素和慶大霉素。另外氯霉素、紅霉素、四環(huán)素〔或多西環(huán)素、或米諾環(huán)素〕及利福平可用于VRE株。Vancomycinresistantenterococciiscurrentlynoeffectivetreatment,buttheVREtopenicillin-sensitivepenicillinandgentamicincombinationtherapycanbeused,ifnotthehigh-levelresistancetopenicillin-resistantavailableaminoglycosidesneomycinandcelebrategreatwhiteAdriamycin.Additionofchloramphenicol,erythromycinandtetracycline(ordoxycycline,orminocycline)andrifampincanbeusedforVREstrains.臨床意義

Clinicalsignificance耐萬古霉55〔三〕病原菌耐藥基因的檢測(cè)Detectionofpathogenresistancegenes更早Earlier確證Confirmed準(zhǔn)確Accurate金標(biāo)準(zhǔn)Goldstandard〔三〕病原菌耐藥基因的檢測(cè)Detectionofpath56第三節(jié)臨床感染常見病原體檢查Checkthecommonpathogensofclinicalinfection

第三節(jié)臨床感染常見病原體檢查Checkthecommo57感染性疾病指各種生物性病原體〔病原微生物、寄生蟲〕寄生人體所引起的傳染性感染疾病和非傳染性感染疾病。Referstoavarietyofinfectiousdiseases,biologicalpathogens(pathogens,parasites)causedbyhumaninfectiousparasiticdiseasesandnon-communicablediseases.感染性疾病指各種生物性病原體〔病原微生物、寄生蟲〕寄生人體所58一、流行病學(xué)和臨床類型Epidemiologyandclinicaltypes一、流行病學(xué)和臨床類型Epidemiologyandc59〔一〕流行病學(xué)Epidemiology新傳染病陸續(xù)被發(fā)現(xiàn)，老傳染病死灰復(fù)燃Beendiscoverednewinfectiousdiseases,theresurgenceofolddiseases多重耐藥株Multi-drugresistantstrains新醫(yī)療技術(shù)的開展Thelaunchingofnewmedicaltechnologies〔一〕流行病學(xué)Epidemiology新傳染病陸續(xù)被發(fā)現(xiàn)，老60〔二〕臨床類型Clinicaltypes臨床常見病原體為：細(xì)菌、真菌、病毒、支原體、衣原體、螺旋體、立克次體以及寄生蟲等。Commonclinicalpathogens:bacteria,fungi,viruses,mycoplasma,chlamydia,spirochetes,rickettsia,andparasites.〔二〕臨床類型Clinicaltypes臨床常見病原體為：61二、檢查工程和臨床應(yīng)用inspectionitemsandclinicalapplications二、檢查工程和臨床應(yīng)用inspectionitems62〔一〕細(xì)菌感染檢查工程的選擇

Theselectionofcheckingprojectsonbacterialinfection普通細(xì)菌：常采用直接鏡檢、別離培養(yǎng)與鑒定、抗原檢測(cè)。Commonbacteria:directmicroscopicexaminationisoftenused,Isolationandidentificationofantigen.病原體抗體檢測(cè)。Antibodydetectionofpathogens難培養(yǎng)或培養(yǎng)要求高的細(xì)菌：綜合運(yùn)用免疫學(xué)或分子生物學(xué)檢測(cè)，如結(jié)核分枝桿菌感染以及幽門螺桿菌感染等。Difficultforhighcultureorbacterialculture:theintegrateduseofimmunologyormolecularbiology,suchasMycobacteriumtuberculosisandHelicobacterpyloriinfection.細(xì)菌培養(yǎng)是最重要確實(shí)診方法。Bacterialcultureisthemostimportantdiagnosticmethod.〔一〕細(xì)菌感染檢查工程的選擇

Theselectiono63〔二〕實(shí)驗(yàn)結(jié)果分析和臨床應(yīng)用Analysisandclinicalapplicationofexperimentalresults顯微鏡檢查或別離培養(yǎng)的陰性結(jié)果不能完全排除感染；Microscopyorisolationandcultureofnegativeresultscannotbecompletelyruledoutinfection共同抗原引起的穿插反響；Antigeniccross-reactivitycausedbythecommon核酸檢測(cè)的假陽(yáng)性；False-positivenucleicaciddetection血清學(xué)的動(dòng)態(tài)檢測(cè)。Dynamicdetectionofserological〔二〕實(shí)驗(yàn)結(jié)果分析和臨床應(yīng)用Analysisandcli64第六節(jié)醫(yī)院感染常見病原體檢測(cè)Detectionofcommonpathogensofnosocomialinfection

第六節(jié)醫(yī)院感染常見病原體檢測(cè)Detectionofco65醫(yī)院感染〔nosocomialinfection〕：又稱醫(yī)院獲得性感染〔hospitalacquiredinfection〕，指患者在入院時(shí)不存在，也不處于潛伏期，而在醫(yī)院內(nèi)發(fā)生的感染，包括醫(yī)院內(nèi)獲得的而在出院后發(fā)病的感染。廣義的醫(yī)院感染對(duì)象不僅指住院患者，還包括門診病人、醫(yī)院職工、探視人員與陪護(hù)人員等。Nosocomialinfection(nosocomialinfection):alsoknownashospital-acquiredinfection(hospitalacquiredinfection),thatdoesnotexistonadmissionofpatients,norintheincubationperiod,whileinthehospitalinfectionsoccurred,includingthehospitalafterdischargefromhospitalacquireddiseaseinInfection.Generalizedinfection,notonlybymeansofhospitalinpatients,includingoutpatient,hospitalstaff,visitingofficersandescortpersonnel.醫(yī)院感染〔nosocomialinfection〕：又稱醫(yī)66一、流行病學(xué)和臨床類型Epidemiologyandclinicaltypes一、流行病學(xué)和臨床類型Epidemiologyandc67〔一〕流行病學(xué)Epidemiology〔一〕流行病學(xué)Epidemiology68病原學(xué)

Pathogen細(xì)菌是最常見的病原體。Bacterialisthemostcommonpathogens.變化趨勢(shì)：革蘭氏陰性桿菌比例在增加，革蘭氏陽(yáng)性球菌比例在減少。Trend:increasingtheproportionofGram-negativebacteria,Gram-positivecocciratiodecreased.罕見細(xì)菌變成流行株：陰溝腸桿菌、洋蔥假單孢菌，軍團(tuán)菌等Strainsofbacteriabecomerare:Enterobactercloacae,Pseudomonasonion,Armymonocytogenes其它：病毒、真菌等Other:viruses,fungi,etc病原學(xué)

Pathogen細(xì)菌是最常見的病原體。692.感染源

Sourceofinfection住院病人、醫(yī)院職工、探視人員、陪伴人員；Patients,hospitalstaff,visitingstaff,hiscompanion;醫(yī)院環(huán)境、未徹底消毒的醫(yī)療器械、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，如腫瘤、糖尿病以及肝硬化患者等Patientswithchronicdiseasessuchascancer,diabetesandlivercirrhosis,etc.

免疫抑制劑及激素的使用者Immunosuppressivedrugsandhormoneusers

化療及放療患者Chemotherapyandradiotherapyinpatients

手術(shù)及麻醉患者Surgeryandanesthesiainpatients

使用介入性診斷與治療的患者Useofinterventionaldiagnosisandtreatmentofpatients

廣譜抗生素使用者Broad-spectrumantibioticusers高危人群

High-riskgroups慢性疾病患者，如腫71〔二〕常見臨床類型

Commonclinicaltypes1下呼吸道感染：最常見Lowerrespiratorytractinfection:themostcommon2尿路感染UrinaryTractInfection3手術(shù)切口感染：外源性和內(nèi)源性Surgicalwoundinfection:exogenousandendogenous4胃腸道感染Gastrointestinalinfections5血液感染Bloodinfection6皮膚和軟組織感染Skinandsofttissueinfections〔二〕常見臨床類型

Commonclinicaltype72二、檢查工程和臨床應(yīng)用inspectionitemsandclinicalapplications二、檢查工程和臨床應(yīng)用inspectionitemsa73〔一〕醫(yī)院感染病原體檢查工程和臨床應(yīng)用inspectionitemsandclinicalapplicationsforHospitalInfection〔一〕醫(yī)院感染病原體檢查工程和臨床應(yīng)用74根本原那么與病原體感染類似，但更應(yīng)注意正常菌群的污染以及條件致病菌在醫(yī)院感染中的地位。Similartothebasicprinciplesandpathogeninfection,butmoreshouldpayattentiontothepollutionofthenormalfloraandopportunisticpathogensinthehospitalinfectionstatus.根本原那么與病原體感染類似，但更應(yīng)注意正常菌群的污染以及條件75標(biāo)本采集和送檢根本原那么

Thebasicprinciplesofspecimencollectionandsubmission及時(shí)采集標(biāo)本Collectspecimensinatimelymanner嚴(yán)格無菌操作Strictaseptic立即送出Immediatelysent在抗菌藥物使用前Beforetheuseofantibiotics適宜的容器Appropriatecontainer注明來源和目的Indicatethesourceanddestination定量或半定量培養(yǎng)Quantitativeorsemi-quantitativeculture分級(jí)報(bào)告和限時(shí)報(bào)告Reportsandlimitgradingreport標(biāo)本采集和送檢根本原那么

Thebasicprincip762.涂片鏡檢

Smearmicroscopy痰標(biāo)本Sputum2.涂片鏡檢

Smearmicroscopy痰標(biāo)本773.別離培養(yǎng)鑒定法

Isolationandcultureidentificationmethod尿路感染UrinaryTractInfection手術(shù)切口感染Surgicalwoundinfection大便培養(yǎng)Stoolculture血培養(yǎng)Bloodculture3.別離培養(yǎng)鑒定法

Isolationandcultu78〔二〕醫(yī)院環(huán)境中細(xì)菌污染的監(jiān)測(cè)和消毒滅菌效果的監(jiān)測(cè)Bacterialcontaminationinthehospitalenvironmentmonitoringandthemonitoringofsterilizationresults〔二〕醫(yī)院環(huán)境中細(xì)菌污染的監(jiān)測(cè)和消毒滅菌效果的監(jiān)測(cè)79定期對(duì)醫(yī)院環(huán)境進(jìn)展病原體監(jiān)測(cè)：空氣、物體外表、醫(yī)務(wù)人員手部；Regularmonitoringofpathogensinhospitalenvironment:air,surface,themedicalstaffofhand;定期對(duì)醫(yī)療器械的消毒效果進(jìn)展監(jiān)測(cè)：高壓蒸氣、紫外線、化學(xué)消毒劑。Regulardisinfectionofmedicaldevicestomonitor:high-pressuresteam,UVlight,chemicaldisinfectants.定期對(duì)醫(yī)院環(huán)境進(jìn)展病原體監(jiān)測(cè)：空氣、物體外表、醫(yī)務(wù)人員手部；80Thanks！Thanks！81臨床常見病原體檢測(cè)

Examinationofclinicalcommonpathogens呼吸內(nèi)科熊維寧DepartmentofRespiratoryMedicineXiong,Weining臨床常見病原體檢測(cè)

Examinationofclin82目的

Objective確定感染的發(fā)生和性質(zhì)，及早明確診斷；Determinetheoccuringandnatureofinfection,andmakediagnosisearly;盡早選擇適當(dāng)?shù)闹委煼桨?；Selecttheappropriatetreatmentassoonaspossible;采取有效的預(yù)防措施，防止感染可能廣泛傳播所造成的危害。Takeeffectivepreventionmeasurestopreventinfectiontocausewidespreaddamage.目的

Objective確定感染的發(fā)生和性質(zhì)，及早明確診斷；83根本程序

BasicProcedures正確采集和運(yùn)送標(biāo)本；Correctcollectionandtransportationofspecimens初步診斷：直接鏡檢，免疫學(xué)和分子生物學(xué)檢測(cè)；Initialdiagnosis:directmicroscopy,immunologyandmolecularbiologyexamination;確定診斷：病原體的別離、鑒定及藥敏實(shí)驗(yàn)，報(bào)告結(jié)果；Determineddiagnosis:pathogenisolation,identificationandsensitivitytest,reportingresults;合理用藥Appropriatedruguse根本程序

BasicProcedures正確采集和運(yùn)送標(biāo)本84第一節(jié)

SectionOne標(biāo)本的采集運(yùn)送、實(shí)驗(yàn)室評(píng)價(jià)和檢查方法Transportationandcollectionofspecimen,laboratoryevaluationandexaminationmethods第一節(jié)

SectionOne標(biāo)本的采集運(yùn)送、實(shí)驗(yàn)室評(píng)價(jià)和檢85一、標(biāo)本采集和運(yùn)送One.CollectionandtransportationofSpecimen一、標(biāo)本采集和運(yùn)送86根本原那么

Basicprinciples根據(jù)病史與臨床表現(xiàn)確定標(biāo)本采集的時(shí)間、部位、種類和數(shù)量，盡量采集病變明顯部位標(biāo)本；Accordingtohistoryandclinicalmanifestationstodeterminethecollectiontime,location,typeandquantityofspecimens,collectspecimensatsignificantpartsoflesions;無菌操作，防止污染；Useaseptictechniquetopreventcontamination;采取適宜的方式進(jìn)展儲(chǔ)存，盡快送檢；Appropriatewaytostore,assoonaspossibleforexamination;要視所有標(biāo)本為傳染品，高度危險(xiǎn)性的標(biāo)本要有明顯標(biāo)識(shí)，急癥或危重患者標(biāo)本要特別注明。Toviewallthespecimensforinfectious,highriskspecimensshouldbeclearlymarked,acuteorcriticallyillpatientspecimensshouldbespecified.根本原那么

Basicprinciples根據(jù)病史與臨床表87臨床常見病原體檢測(cè)英文版教學(xué)課件88臨床常見病原體檢測(cè)英文版教學(xué)課件89〔二〕尿液

(Two)Urine無菌采集中段尿；Sterilecollectionofmid-portionurine;如考慮厭氧菌感染，采取膀胱穿刺法采集標(biāo)本，無菌厭氧小瓶運(yùn)送；Ifconsiderationofanaerobicinfections,applybladderpuncturetocollectspecimens,transportthemwithsterileanaerobicvial;排尿困難者考慮導(dǎo)尿采集標(biāo)本。Applycatheterizationtocollectspecimensfordysuriapatients.〔二〕尿液

(Two)Urine無菌采集中段尿；90〔三〕糞便

(Three)Stool挑取膿、血或粘液局部于清潔容器中送檢；Pickpus,bloodormucusfromstollintocleancontainersforexamination;排便困難者或嬰兒采用直腸拭子采集，置于有保存液的試管內(nèi)送檢；Applyrectalswabforinfantsordifficultdefecation,placeitwithpreservationsolutioninthetesttubeforexamination;疑心霍亂弧菌感染引起的腹瀉，將標(biāo)本置于堿性蛋白胨水或卡-布〔Cary-Blair〕運(yùn)送培養(yǎng)液送檢；SuspectedinfectionscausedbyVibriocholerae,thespecimenswereplacedinalkalinepeptonewaterorcard-cloth(Cary-Blair)transportmediumforexamination;傳染性腹瀉應(yīng)連續(xù)送檢3次。Continuousexamination3timesforinfectiousdiarrhea.〔三〕糞便

(Three)Stool挑取膿、血或粘液局部于91〔四〕呼吸道標(biāo)本

(Four)Respiratoryspecimens類型：鼻咽拭子，痰和經(jīng)氣管采集的標(biāo)本；Types:nasopharyngealswab,sputum,andspecimenscollectedthroughtracheal上呼吸道存在正常菌群，在采集標(biāo)本與結(jié)果分析時(shí)應(yīng)予考慮。Thereisnormalflorainupperrespiratorytract,thespecimenscollectionandresultsanalysisshouldbeconsidered.〔四〕呼吸道標(biāo)本

(Four)Respiratorysp92〔五〕腦脊液與其他無菌體液

(Five)Cerebrospinalfluidandothersterilebodyfluids腦脊液應(yīng)立即保溫送檢或床邊接種；CSFshouldbetransportedimmediatelywithinsulationorculturedbedside；胸腔積液、腹腔積液和心包積液等應(yīng)采集較大量標(biāo)本送檢，離心后再接種培養(yǎng)。Pleuraleffusion,ascitesandpericardialeffusionwithalargevolumeshouldbecollected,centrifugatedandthencultured.〔五〕腦脊液與其他無菌體液

(Five)Cerebrosp93〔六〕眼、耳部標(biāo)本

(Six)Eye,earspecimens拭子采樣Swabsamples〔六〕眼、耳部標(biāo)本

(Six)Eye,earspeci94〔七〕泌尿生殖道標(biāo)本

(Seven)Urogenitalspecimens男性：無菌采集尿道口分泌物或前列腺液；Male:urethraorprostaticfluidsecretionswithsterilecollection;女性：無菌采集陰道或?qū)m頸分泌物。Female:vaginalorcervicalsecretionscollectedaseptically.〔七〕泌尿生殖道標(biāo)本

(Seven)Urogenital95〔八〕創(chuàng)傷、組織和膿腫標(biāo)本

(Eight)Trauma,organization,andabscessspecimens對(duì)損傷范圍較大的創(chuàng)傷，從不同部位采集多份標(biāo)本；Onawiderrangeoftraumainjuries,specimenscollectedfrommanydifferentparts;采集部位應(yīng)首先去除污物、消毒皮膚；Firstlyremovedirt,disinfecttheskinaroundcollectionsite;標(biāo)本較少那么需參加無菌生理鹽水以防枯燥；Fewersamplesmustbeaddedtosterilesalinetopreventdrying;開放性膿腫及膿性分泌物：用無菌棉纖采取膿液或病灶深局部泌物；Openabscessandpurulentsecretions:collectdeepdischargepusorlesionswithsterilecottonfibers;封閉性膿腫用注射器抽取；Collectspecimeninclosedabscesswithasyringe;疑心厭氧菌感染，應(yīng)隔絕空氣采集。Suspectedanaerobicinfections,collectionshouldbeisolatedfromair.〔八〕創(chuàng)傷、組織和膿腫標(biāo)本

(Eight)Trauma,96〔九〕血清

(Nine)Serum用于檢測(cè)特異性抗體；Forthedetectionofspecificantibodies;血液自然凝固后吸取血清，滅活補(bǔ)體。Drawserumafternaturalcoagulationofblood,theninactivatecomplement.〔九〕血清

(Nine)Serum用于檢測(cè)特異性抗體；97二、標(biāo)本的實(shí)驗(yàn)室質(zhì)量評(píng)估標(biāo)準(zhǔn)

Two.Standardoflaboratoryqualityassessmentofspecimens二、標(biāo)本的實(shí)驗(yàn)室質(zhì)量評(píng)估標(biāo)準(zhǔn)

Two.Standardo98檢驗(yàn)申請(qǐng)單的根本內(nèi)容

ThebasiccontentofanapplicationforexaminationT