医学微生物课件.ppt

1、v掌握：掌握：1.结核分枝杆菌的形态染色、培养特性、抵抗力、结核分枝杆菌的形态染色、培养特性、抵抗力、致病机理、结核菌素试验的原理与用途、微致病机理、结核菌素试验的原理与用途、微生物学检查法、卡介苗预防；生物学检查法、卡介苗预防；2.麻风分枝杆菌的形态染色、致病特点。麻风分枝杆菌的形态染色、致病特点。v熟悉：结核分枝杆菌的免疫与变态反应关系。熟悉：结核分枝杆菌的免疫与变态反应关系。2020/11/32v aerobic,v inability to be Gram-stainedv Lipid-rich cell wall,especially mycolic acid分枝菌酸，分枝菌酸，vr

2、esist decolorization with acid-alcohol after stained with carbolic fuchsinvacid-fast stain,acid-fast rods.2020/11/33细长略弯的杆菌，有分枝生长趋势；细长略弯的杆菌，有分枝生长趋势；细胞壁含大量脂质、不易着色，染色细胞壁含大量脂质、不易着色，染色后可抵抗盐酸乙醇脱色，后可抵抗盐酸乙醇脱色，抗酸杆菌抗酸杆菌。2020/11/341.1.M.Tuberculosis bacillus complexM.Tuberculosis bacillus complex 结核分枝杆菌复合群结核分

3、枝杆菌复合群 生长缓慢，包括人和牛结核分枝杆菌生长缓慢，包括人和牛结核分枝杆菌2.Nontuberculous mycobacteria2.Nontuberculous mycobacteria 非结核分枝杆菌非结核分枝杆菌3.M.leprae3.M.leprae麻风分枝杆菌麻风分枝杆菌分枝杆菌的分类分枝杆菌的分类classifyclassify2020/11/35Acid Fast StainAcid Fast Stain(Ziehl-Neelsen stain)(Ziehl-Neelsen stain)vStain the slide with carbol fuchsin 石炭酸复石炭酸

4、复红红(a red dye)vDecolorize it with 3%hydrochloric acid盐酸酒盐酸酒精精(acid-alcohol).vcounter-stain with methylene blue.美蓝美蓝 The Lipid especially mycolic acid can resist to decolorization with acid-alcohol after stained with carbol fuchsin2020/11/36抗酸染色阳性抗酸染色阳性抗酸染色阴性抗酸染色阴性固定固定5 5石炭酸复红石炭酸复红3 3盐酸酒精盐酸酒精美蓝美蓝2020

5、/11/37Mycobacterium stains red other bacterium and the background stains blue.Acid Fastness Stain(Ziehl-Neelsen stain)2020/11/382020/11/392020/11/310 Pathogens1.Mycobacterium tuberculosis2.Mycobacterium leprae2020/11/311Mycobacterium tuberculosisacid-fast stain2020/11/3122020/11/313v9岁，男孩，因发热，咳嗽咳痰入院

6、，胸部岁，男孩，因发热，咳嗽咳痰入院，胸部X光检查可见肺部有哑铃状阴影（原发综合征光检查可见肺部有哑铃状阴影（原发综合征表现）：表现）：v（1）你认为该儿童得的是什么病？）你认为该儿童得的是什么病？v（2）为了明确诊断，你会对他做哪些微生物）为了明确诊断，你会对他做哪些微生物学检查？学检查？2020/11/314第 一 节 结 核 分 枝 杆 菌第 一 节 结 核 分 枝 杆 菌 Mycobacterium tuberculosis（tubercle bacilli）。2020/11/315Morphologyv thin and slightly cur rodsv acid-fast ba

7、cilli2020/11/316形态染色：形态染色：细长略弯的杆菌，常聚集成团。细长略弯的杆菌，常聚集成团。2020/11/317 抗酸染色法：分枝杆菌呈抗酸染色法：分枝杆菌呈红色红色，其它菌及背景为蓝色。其它菌及背景为蓝色。2020/11/318Mycobacterium tuberculosis2020/11/3191.obligate aerobic.mycodermmycoderm菌膜菌膜2.nutrition are high,Lowenstein-Jensen medium 3.It grows very slow with a generation time of 18 hour

8、s.colony,3-4 weeks4.The rough colonies granular,dull-yellow,dryCulture 2020/11/320培养特性：培养特性：1 1、专性需氧。、专性需氧。液体培养基中形成液体培养基中形成mycodermmycoderm菌膜，菌膜，索状生长。索状生长。3 3、营养要求高，罗氏培养基。、营养要求高，罗氏培养基。4 4、生长缓慢。、生长缓慢。5 5、菌落呈颗粒、结节或花菜状，、菌落呈颗粒、结节或花菜状，乳白或米黄色，不透明。乳白或米黄色，不透明。2020/11/3212020/11/3222020/11/3232020/11/3242020

9、/11/325vResistant dry (sputum)vRelatively resistant to acids and alkalivResistant to the main antimycobacterial drug drug resistancevSensitive to ultra-violet light,heat,and alcohol 2020/11/326抵抗力：抵抗力：干燥干燥 抵抗力强抵抗力强 酸、碱酸、碱 乙醇、湿热、紫外线乙醇、湿热、紫外线 药物敏感，但易耐药药物敏感，但易耐药敏感敏感有抵抗力有抵抗力2020/11/327vShape VariationvC

10、olony VariationvVirulent VariationvDrug resistance Variation2020/11/328A.Constituents of Tubercle Bacilliv no toxinsvsurvive and multiplies within cells2020/11/329二、致病性二、致病性（一）致病物质（一）致病物质1 1、脂质：、脂质：(1)(1)Cord factor索状因子：抑制白细胞索状因子：抑制白细胞游走，与慢性肉芽肿形成有关。游走，与慢性肉芽肿形成有关。(2)(2)Phosphatides磷脂：促使单核细磷脂：促使单核细胞增生

11、胞增生TbTb结节形成，干酪样坏死。结节形成，干酪样坏死。2020/11/330(3)(3)Wax D蜡质蜡质D D：有佐剂作用，激：有佐剂作用，激发机体产生迟发型变态反应。发机体产生迟发型变态反应。(4)(4)Sulfatide硫酸脑苷脂：使硫酸脑苷脂：使TbTb杆菌杆菌在吞噬细胞中大量存活。在吞噬细胞中大量存活。2020/11/331 2.Several proteins When combined with wax D,elicit delayed hypersensitivity(tuberculin reaction)3.Polysaccharides 2020/11/3322 2、

12、蛋白质：、蛋白质：TbTb菌素蜡质菌素蜡质D D -迟发型超敏反应迟发型超敏反应3 3、多糖、多糖2020/11/333Several complex lipidsvPhosphatides 磷脂磷脂:play a role in caseous necrosisvCord factor：Correlated with virulence of the organism Inhibits migration of leukocytes,causes chronic granulomas,can serve as adjuvantvSulfatide硫酸脑苷脂硫酸脑苷脂 suppress pha

13、gosome combine with lysosomevWax D:adjuvant,used to enhance the immune response2020/11/334Diseases tuberculosis approximately one third of the worlds population is infected2020/11/335Pathogenesis primary infection1)lung infection secondary infection 2)Out lung infection :intestines肠、肾肠、肾kidney、bone、

14、brain,joint,and so on.2020/11/336vOccur in the lungsvno immunity(children)vGhon complex:(Ghon focus)（survive and multiplies within macrophage）initial lesion and enlarged hilar lymph nodesv90%heal but a few bicillin in lung vSome bacilli diffusion 2020/11/3371 1、肺部感染、肺部感染：肺结核：肺结核最多见最多见（1 1）原发感染）原发感染

15、TbTb杆菌杆菌肺泡肺泡大量繁殖大量繁殖肺泡肺泡内炎症内炎症(原发灶原发灶)Tb)Tb杆菌通过淋巴管杆菌通过淋巴管到肺门淋巴组织到肺门淋巴组织肺门淋巴结肿大肺门淋巴结肿大(原发综合征原发综合征)自愈自愈(钙化、纤维钙化、纤维化化)（约（约90%90%）。少数免疫力低下者，）。少数免疫力低下者，经血流播散经血流播散 全身结核病全身结核病2020/11/338vAdultsvectogenous infection or or autogenous infection vFocus of infection are limitedvCavity formation2020/11/339（2 2）原

16、发后感染：）原发后感染：可为外源性或内源性感染，肺部可为外源性或内源性感染，肺部多见，病灶局限，一般不累及邻近多见，病灶局限，一般不累及邻近的淋巴结。易形成空洞。的淋巴结。易形成空洞。2020/11/340Cavity formation2020/11/341v原发结核原发结核：外源性感染：外源性感染多发生于儿童（无免疫力）多发生于儿童（无免疫力）原发综合征原发综合征 (肺门淋巴肿大等肺门淋巴肿大等)形成结核结节（即结核肉芽肿），形成结核结节（即结核肉芽肿），少数病灶扩散，少数病灶扩散，引起全身引起全身TBTB转归转归v原发后结核原发后结核：外或内源性感染：外或内源性感染多见成人多见成人病灶局

17、限病灶局限 引起局部引起局部TBTB、开放性、开放性TBTB（反应剧烈，（反应剧烈，干酪性坏死、空洞）干酪性坏死、空洞）2020/11/342肺结核病的主要症状有：肺结核病的主要症状有：coughcough咳嗽、咳痰咳嗽、咳痰spit bloodspit blood咯血、咯血、chest painchest pain胸痛胸痛fever in afternoonfever in afternoon、fatiguefatigue疲乏无力、疲乏无力、anorexiaanorexia食欲减退食欲减退 、lose fleshlose flesh消瘦、消瘦、night sweatsnight sweats

18、夜间盗汗、夜间盗汗、2020/11/343Resistance mainly by cellular immunityHigh rate of infection,but low morbidity.2020/11/344phenomenon2020/11/345 not diffusion,heal rapidly 有毒TB菌Diffusion郭霍现象郭霍现象2020/11/346(二二)免疫与变态反应：免疫与变态反应：郭霍现象：郭霍现象：原发感染特点：原发感染特点：初次感染结核杆菌，初次感染结核杆菌，10-1410-14天局部出现溃疡，天局部出现溃疡，深深,不易不易愈合；附近淋巴结愈合；附

19、近淋巴结肿大肿大，细菌，细菌易扩散易扩散。原发后感染特点：原发后感染特点：再次感染结核杆菌，再次感染结核杆菌，1-21-2天局部出现溃疡，天局部出现溃疡，浅浅,易易愈合；附近淋巴结愈合；附近淋巴结不肿大不肿大，细菌，细菌不扩散不扩散。2020/11/347v初次注射初次注射 局部反应小局部反应小 全身扩散全身扩散 无免疫无超敏反应无免疫无超敏反应v再次注射再次注射 局部反应大局部反应大 不扩散不扩散 有免疫有超敏反应有免疫有超敏反应2020/11/348vDue to a delayed hypersensitivity reactionvOT PPD:antigen,contain 5 tu

20、berculin unitsvEvaluated by measuring the diameter of the induration surrounding the test site 2020/11/349(三三)结核菌素试验：结核菌素试验：1 1、原理：原理：应用结核菌素进行皮肤试验测应用结核菌素进行皮肤试验测定机体对结核分枝杆菌是否引起超定机体对结核分枝杆菌是否引起超敏反应的一种试验。敏反应的一种试验。2020/11/350Mantoux method When the Mantoux skin test is performed,a needle is injected into

21、the upper skin layer of the patients arm.The arm is examined 48 to 72 hours after the tuberculin injection in order to evaluate the reaction on the patients skin.Any swelling that can be felt around the site of the injection,also known as induration,is measured.The diagnosis of TB infection depends

22、on the size of the measured induration and the patients individual risk factors.2020/11/3512 2、结核菌素：、结核菌素：旧结核菌素旧结核菌素(OT)(OT)：结核菌菌体蛋白：结核菌菌体蛋白,纯蛋白衍化物纯蛋白衍化物(PPD)(PPD)3 3、试验方法：、试验方法：常规试验取常规试验取PPD5uPPD5u注射两前臂皮内，注射两前臂皮内，48724872小时后观察结果小时后观察结果2020/11/352vpositive：Induration 5 mm caused by a delayed hyperse

23、nsitivity response Indicates previous infection but not necessarily active diseaseresult2020/11/353vstrong positive(+):Induration 15 mm,diagnosis of infant tuberculosis.vnegative(-)Induration 5mm or no Induration:no infection and no immunity2020/11/354negative(-)：vIn the early days of infectionvThe

24、agedvOverwhelming tuberculosis or other diseasevAIDS2020/11/355 4 4、结果观察及意义、结果观察及意义(1)(1)阳性：局部红肿、硬结阳性：局部红肿、硬结5mm,5mm,感染感染免疫免疫变态反应。变态反应。(2)(2)强阳性：局部红肿、硬结强阳性：局部红肿、硬结15mm15mm，有诊断意义。，有诊断意义。(3)(3)阴性：局部无反应或硬结阴性：局部无反应或硬结5mm5mm未感染过结核杆菌。未感染过结核杆菌。2020/11/356 a a、感染初期。、感染初期。b b、老年人。、老年人。c c、严重结核病患者或正患其他、严重结核病患

25、者或正患其他 传染病。传染病。d d、获得性细胞免疫低下。、获得性细胞免疫低下。阴性者应考虑几种情况：阴性者应考虑几种情况：2020/11/3572020/11/3582020/11/359Laboratory diagnosis1.Specimen:sputum,pus,CSF,urine,etc.2.Microscopic examination:Ziehl-Neelsen stain3.Concentration:4%NaOH-3%HCL;6%H2SO44.Culture:solid culture(2-4 weeks 37);liquid culture(1-2 weeks)5.Ani

26、mal inoculation:guinea pig6.quick Diagnosis:PCR 2020/11/360微生物学检查微生物学检查1 1、标本：、标本：根据感染部位选材，根据感染部位选材，某些部位标本应先集菌，如离心或消化。某些部位标本应先集菌，如离心或消化。2 2、直接涂片镜检：抗酸染色。、直接涂片镜检：抗酸染色。3 3、分离培养。、分离培养。4 4、动物试验。、动物试验。5 5、快速诊断。、快速诊断。2020/11/361PreventionVaccineBCG vaccination for new infants 2020/11/3622020/11/3632020/11/

27、364v结核分枝杆菌的形态染色、培养特点。结核分枝杆菌的形态染色、培养特点。v结核杆菌的免疫特点？结核杆菌的免疫特点？v什么叫结核菌素试验？其原理是什么？结果什么叫结核菌素试验？其原理是什么？结果有何意义？有何意义？v试述结核杆菌的微生物学检查方法。试述结核杆菌的微生物学检查方法。2020/11/3652020/11/366HANSENS DISEASE(Leprosy)caused by M.lepraevHansens disease is a chronic,slowly progressive granulomatous disease involving ectodermally d

28、erived tissue such as the skin and peripheral nerves.The disease is usually limited to the cooler parts of the body such as the skin,nose and upper respiratory tract.It rarely affects internal organs such as the brain,liver,spleen,kidneys,and bones.vIt has a specific predilection for peripheral nerv

29、es.2020/11/367 aerobic,acid-fast rods cant be cultured in vitro 2020/11/368 麻风分枝杆菌麻风分枝杆菌生物学性状：生物学性状：与结核菌相似，常呈束状排列，与结核菌相似，常呈束状排列，人工培养未成功。人工培养未成功。2020/11/369Leprosy bacilliacid-fast stain2020/11/370 habitat:human skin and superficial nerves transmission:by aerosol or prolonged contact with patients wi

30、th lepromatous leprosy2020/11/3712020/11/372v传播方式传播方式呼吸道呼吸道也可通过接触传染也可通过接触传染v发病慢发病慢,病程长病程长v麻风的免疫主要是靠细胞免疫麻风的免疫主要是靠细胞免疫v细菌主要侵犯外周神经、皮肤细菌主要侵犯外周神经、皮肤2020/11/373vlesions:in the cooler parts of the bodyvReplicate intracellularly,typically within skin histiocytes,endothelial cells and the Schwann cells of ne

31、rves 2020/11/3742 types:2 forms:2020/11/375临床分为临床分为 两型两型(瘤型、结核样型瘤型、结核样型)两类两类(界线类、未定类界线类、未定类)。2020/11/376lepromatous leprosy:瘤型麻风瘤型麻风1.the cell-mediated response to Leprosy bacilli is poor 2.large numbers of organisms appear in the lesions and blood(bacteremia)(powerful infectious)3.involving the ski

32、n and mucous membranes4.multiple nodular skin lesions,leonine facies2020/11/3772020/11/378The face of a patient with active,neglected nodulous lepromatous leprosy.With treatment,all nodules could be reversed.WHO/TDR/McDougall2020/11/3792020/11/380Treated lepromatous leprosy.The nodularity of the ski

33、n has resolved on treatment but the absence of eyebrows and the nasal collapse remain.2020/11/3812020/11/382:1.the cell-mediated response(normal)limit the growth of the organism2.Very few acid-fast bacilli are seen (lower infectious）3.involving the skin and peripheral nerves.4.macular skin lesions t

34、hickened superficial nerves significant anesthesia of skin 2020/11/383v The organism also has a strong affinity for nerves.Deformity due to nerve damage with its consequent ulcers and resorption of bone.Such deformities can be worsened by careless use of the hands.WHO/TDR2020/11/3842020/11/3852020/11/3862020/11/3872020/11/388Thank You世界触手可及世界触手可及携手共进，齐创精品工程携手共进，齐创精品工程