1. Widal’s Reaction Results

2. 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.

3. H 抗体与 O 抗体的性质及其消长的意义 O 抗体 IgM 出现较早 维持时间短 H 抗体 IgG 出现较晚 维持时间长 伤寒的可 能性很大 高 高 高 低 感染早期 O 凝集的交 叉反应 低 高 预防接种 非特异性回忆 反应 抗体类型 体内出 现情况 与正常值比较 Analysis

4. Observation 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.

5. Experiment five Isolation and Identification of influenza virus

6. 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

7. 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 morphology/immune electron microscopy  Light microscopy for histological appearance - e.g. inclusion bodies  Antigen detection with immunofluorescence, ELISA etc.  Molecular techniques for the direct detection of viral genomes

8. 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

9. 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.

10. 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.

11. 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

12.  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. 流感病毒的分离鉴定

13. Hemagglutination inhibition test complement fixation test Inoculate to fresh embryos after two such passages Treated with penicillin and streptomycin Embryonated egg inoculation (amniotic or allantoic route) Harvest amniotic and allantoic fluids Hemagglutination test the result is negative +- Throat washings or gargling Patient’s serum 血凝抑制实验

14. 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.

15. Materials well No. (ml) 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 弃 Dilution 1:10 1:20 1:40 1:80 1:160 1:3201:6401:12801:2560 con 0.5%RBC 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

16. 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.

18. 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.

19. Materials well No. (ml) 1 2 3 4 5 6 7 8 9 10 Saline 0.9 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0. 5 Serum 0.1 0.25 0.25 0.25 0.25 0.25 0.25 弃 0.25 - 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 - - 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 - - - + ++ +++ ++++ ++++ - - 弃

20. 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.