Survey
* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project
* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project
Widal’s Reaction Results Widal’s Reaction Definition: a specific reaction consisting in agglutination of typhoid bacilli or paratyphoid bacilli when mixed with serum from a patient having typhoid fever or other salmonella infection and constituting a test for the disease . Usage: to assist to diagnosis typhoid fever or other salmonella infection. Analysis H抗体与O抗体的性质及其消长的意义 抗体类型 体内出 现情况 与正常值比较 O抗体 IgM 出现较早 维持时间短 高 高 低 H抗体 IgG 出现较晚 维持时间长 高 低 高 伤寒的可 能性很大 感染早期 预防接种 O凝集的交 非特异性回忆 叉反应 反应 Observation: 1. Control tube: no agglutination (-) 2. Lowest titer tube: absolutely agglutination (++++) 3. Other tubes: three quarters agglutination(+++) half agglutination (++) one quarter agglutination (+) no agglutination (-) Judgment: Agglutination titer: the highest dilution of serum which appears (++) bacteria agglutination. Experiment five Isolation and Identification of influenza virus 病毒感染的微生物学检查法 In general, diagnostic tests can be grouped into 3 categories: direct detection indirect examination (virus isolation) serological methods Direct detection In direct examination, the clinical specimen is examined directly for 病毒颗粒、病毒抗原或核酸 Electron Microscopy electron microscopy morphology/immune Light microscopy for histological appearance e.g. inclusion bodies Antigen detection with immunofluorescence, ELISA etc. Molecular techniques for the direct detection of viral genomes Indirect examination Inoculating the specimen in cell culture, eggs or animals to cultivate virus: this is called 病毒分离. Cell Culture - cytopathic effect, haemadsorption, confirmation by neutralization, interference immunofluorescence etc. Eggs pocks on CAM - haemagglutination, inclusion bodies Animals disease or death confirmation by neutralization Serological diagnosis A serological diagnosis can be made by the detection of rising titres of antibody between acute and convalescent stages of infection, or the detection of IgM. In general, the majority of common viral infections can be diagnosed by serological methods. 快速诊断 Direct examination methods are often also called rapid diagnostic methods because they can usually give a result either within the same or the next day. With the advent of effective antiviral chemotherapy, rapid diagnostic methods are expected to play an increasingly important role in the diagnosis of viral infections. 流感病毒的分离鉴定 Purpose: To obtain information about microbiological diagnosis methods of influenza virus Procedure Acquiring and managing specimen Isolation and Identification of influenza virus serological diagnosis of influenza virus 流感病毒的分离鉴定 标本采集和处理 Throat washing or gargling are obtained within 3 days after infection Test at once or stored frozen. Centrifuge the washings 15 min at 3000 rpm, Harvest 1 ml of supernatant. Add antibiotic to prevent bacterial contamination. Patient’s serum Throat washings or gargling Treated with penicillin and streptomycin 血凝抑制实验 Embryonated egg inoculation (amniotic or allantoic route) Harvest amniotic and allantoic fluids Hemagglutination test + - Hemagglutination inhibition test Inoculate to fresh embryos complement fixation test after two such passages the result is negative Hemagglutination test, HT MATERIALS Virus: the allantoic fluid that contains influenza virus; physiological saline; 0.5% chicken RBC suspension. 20-well plastic disposable microtiter plates and glass pipettes. Hemagglutination test, HT Materials (ml) well No. 1 2 3 4 5 6 7 8 9 10 Saline 0.45 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 Virus 0.05 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 Dilution 1:10 1:20 1:40 1:80 1:160 1:3201:6401:12801:2560 con 0.5%RBC 弃 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 incubate the plate at room temperature for 30 to 60 min. Check the agglutination. Hemagglutination test (HT) Results Record the hemagglutination as the followings: ++++ : All the RBC had been agglutinated. +++ : 75% of RBC had been agglutinated ++ : 50% of RBC had been agglutinated. + : 25% of RBC had been agglutinated. - : No RBC was agglutinated. Hemagglutination inhibition test (HIT) MATERIALS Patient’s serum: deactivated 30 min at 56℃; influenza virus: 4 HA units; 0.5% chicken RBC suspension; physiological saline 20-well microtiter plates, pipettes and tubes. Materials (ml) well No. 1 Saline 2 0.9 3 4 5 6 7 8 9 10 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0. 5 弃 Serum 0.1 Dilution 1:10 1:20 1:40 1:80 1:160 1:320 1:640 Virus 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 弃 0.25 - - - - - - Mix evenly incubation at 37℃ for 1h 0.5%RBC 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 Mix evenly incubation at 20℃ for 1 h Result - - - + ++ +++ ++++ ++++ - - Hemagglutination inhibition test (HIT) RESULT Assess hemagglutination as above, the hemagglutination inhibition (HI) titer is the reciprocal of the highest dilution of the patient's serum which shows complete inhibition of agglutination.