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1、1,第二节 细菌感染检查,2,检验特点,通过对标本直接显微镜检，病原菌的分离培养鉴定，病原菌的抗原检测，核酸检测和细菌感染后机体免疫应答产物的测定及药物的测定及药物敏感性试验，对细菌感染性疾病可作出及时、准确的病原学诊断。,3,检测程序,4,SpecimenStain of smear Culture and isolation Enriched culture Aerobic cultivation Cultivation withcarbon dioxideAnaerobic cultivation Observation of colonial morphology Picking co
2、lony Stain of smear andmicroscopic detectionSerological Biochemical Antimicrobialidentification identification susceptibility testing Determination of pathogenic bacteria and results reporting Procedures for bacterial detection,5,Gram stained smears of clinical specimens provide rapid and presumptiv
3、e evidence of infections.The need for other direct tests must be considered.These issues will determine whether the specimen requires any pretreatment before inoculation.,革兰染色,抗酸染色,染色标本显微镜检测,6,不染色标本显微镜检测(Wet Mounts)A direct wet mount of fecal material,particularly with liquid or unformed stool,is th
4、e fastest method for detection of motile,7,3.分离培养 培养基,普通琼脂平板血琼脂平板选择鉴别琼脂平板：SS、MacC、TCBS等巧克力琼脂平板厌氧血琼脂平板,8,分离培养Culture and isolation,生长条件气体条件:大气条件、厌氧条件、微需氧条件温度：一般35pH值：7.27.6 少数例外Observation of colonial morphology,9,鉴定identification Biochemical identification Serological identification,生化鉴定糖分解产物试验、蛋白质分
5、解产物试验、碳源利用试验、呼吸酶类试验等。,10,生化鉴定,组合生化试验IMViC,11,鉴定技术 血清学鉴定,利用已知菌种（型）的抗体鉴定标本分离菌的种（型），适用于含较多血清型的细菌。,12,鉴定技术 数码鉴定,集数字、电子信息及自动化分析技术于一体，采用标准化、成品化和配套生化反应试条鉴定细菌，利用属、群、亚种和生物型。,API细菌生化鉴定,14,鉴定技术 自动化鉴定,自动化鉴定：自动细菌培养鉴定仪已广泛应用于微生物学检查。有Automicrobic（AMS）、Autobac（IDX）Microscan等系列。应用最广泛、自动化程度最高、功能齐全的是VITEKAMS系统。,微型全自动血培
6、养仪,全自动细菌鉴定及高级专家系统药敏报告仪 VITEK2,17,鉴定技术 核酸检测,PCR适用于细菌感染诊断，尤其是人工培养不能生长的细菌和营养要求高不易培养的细菌。如麻风杆菌、幽门螺杆菌、嗜肺军团菌、结核分枝杆菌。,18,PCR技术,19,The serologic approach includes(1)demonstration of a fourfold or greater rise in antiviral antibody using acute and convalescent sera and a variety of methods that measure predom
7、inantly immunoglobulin(Ig)antibody and(2)demonstration of virus-specific IgM antibody in a single late acute or recovery phase serum.,适用于经抗生素治疗后或慢性细菌性感染。应用时检测双份血清，效价4倍增高，说明曾有感染。血清学诊断宜用于流行病学调查。,鉴 定技术 抗体检测,20,鉴定技术 细菌毒素检测,1.外毒素检测生物学方法:一般不用免疫血清方法:大量样品筛选核酸和基因诊断技术自动化检测2.内毒素检测,21,细菌感染检查项目的选择和应用,按检出细菌种类、临床诊
8、断和治疗，构成不同组合的检验方法.细菌感染诊断原则:1、医师和检验师密切配合2、重视检验师的密切配合3、重视检验过程的阶段性和连续性4、检验程序需有针对性和综合性,22,Diagnosis of bacterial infections,The identification and differentiation of bacteria principally relies:-on microbial morphology and growth variables on culturing the organisms on non-selective and selective media.C
9、ulture remains the standard for diagnosis of bacterial infection because it is the simplest,the most reliable and the most cost efficient method.Another advantage is the fact that parallel testing of the sensitivity towards antibiotics is possible.,23,n molecular-biological technics that provide rap
10、id identification of infectious agents,both from culture isolates and directly within clinical specimens.-on the detection of the antibody response to the pathogen.The concentration of infection-sepcific antibodies is determined in titers or activity units.,24,In principle,the pathogen should be ide
11、ntified by culture in any acute infection that requires treatment.This,however,is only possible if the pathogen is still detectable in excretions,secretions,blood or tissue at the time when the clinical symptoms occur,e.g.salmonellae in feces in the case of diarrhea or gonococci from endocervical canal in genital tract infection.,
