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Diagnosis, prevention and treatment Xiao-Kui Guo PhD

Diagnosis, prevention and treatment Xiao-Kui Guo PhD

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Page 1: Diagnosis, prevention and treatment Xiao-Kui Guo PhD

Diagnosis, prevention and treatment

Xiao-Kui Guo PhD

Page 2: Diagnosis, prevention and treatment Xiao-Kui Guo PhD

Content

Lab diagnosis of the viral infection Prevention of the viral infection Anti-virus treatment

Page 3: Diagnosis, prevention and treatment Xiao-Kui Guo PhD

Section A

Lab Diagnosis of the viral infection

Page 4: Diagnosis, prevention and treatment Xiao-Kui Guo PhD

Methods of the diagnosis

Diagnostic virology ( 病毒感染的检查方法 )provides evidence for the physician to make right diagnosis, as well as the treatment scheme. virus isolation identification and confirmation( 病毒分离与鉴定 ) viral nucleic acid amplification techniques( 病毒核酸检

测技术 ) viral antigen or specific antibodies detection (病毒抗

原及特异性抗体的检测)

Page 5: Diagnosis, prevention and treatment Xiao-Kui Guo PhD

Specimen Collection and Shipment (标本的采集与送检) A. Samples for Virus Isolation and Viral Nucleic

Acid Amplification 1. Timeliness of Specimen Collection 2. Sample Type 3. Sample shipment 4.Process and Stock Samples

B. Samples for Serology Detection

Page 6: Diagnosis, prevention and treatment Xiao-Kui Guo PhD

Virus Isolation

A. Cell Culture B. Animal Inoculation- inoculating a susceptible host (laboratory) a

nimal with infectious material mice, guinea pigs (豚鼠) , rabbits, monkeys, and ferrets (雪貂)

C. Chick Embryo Inoculation Chorioallantoic membrane ( 绒毛尿囊膜 ) (CAM)

①titration of viruses ②investigation of viral morphology ③identification of viruses

Allantoic sac( 尿囊腔 ) -required large quantities of viruses Amniotic sacs ( 卵黄囊 )- the isolation of various viral agen

ts

Page 7: Diagnosis, prevention and treatment Xiao-Kui Guo PhD

A. Cell Culture 1. Cell culture category

(1) Primary cell culture (2) Diploid cell culture (3) Continuous Cell Culture (4) Lymphocyte culture

2. Virus Identification (1) Characterization of Viral Intracellular Propagation

1) Cytopathogenic Effect (细胞致病作用) (CPE) 2) Hemadsorption phenomenon 3) Viral Interference

(2) Quantification of Viral Infection 1) Plaque-forming Unit (空斑形成单位) (PFU) 2) The Tissue Culture Infectious Dose50 (50% 致死量( LD50 )或 5

0% 组织细胞感染量 ) (TCID50) Assay 3) Virus Morphology and Structure ( 病毒形态与结构 )Observation

3. Viral Antigen Detection

Page 8: Diagnosis, prevention and treatment Xiao-Kui Guo PhD

Directly Detect Viral Antigen

Measure viruses without permissive cell line ( 病毒敏感细胞系 ) A. Immunofluorescent Assay

Immunofluorescent microscopy (荧光显微镜) B. Enzyme-Linked Immunosorbent Assays (ELISA)

Page 9: Diagnosis, prevention and treatment Xiao-Kui Guo PhD

Direct Detection of Viral Antibody

Specific viral antigen to detect antibodies in patients’ serum, including IgM antibodies detection and IgG antibodies measurement

Method: The neutralization (Nt) test The complement fixation (CF) test (补体结合实验) The hemagglutination inhibition (HI) test (血凝抑制实验) The immunofluorescence (IF) test Passive hemagglutination Immunodiffusion( 免疫扩散 )

Page 10: Diagnosis, prevention and treatment Xiao-Kui Guo PhD

Virus Nucleic acid detection A. Nucleic acid hybridization

technique Southern blotting(Southern 杂

交 ) Northern blotting(Northern 杂

交 ) Detected through autoradiogra

phy( 放射自显影技术 ) or by the biotin-avidin system ( 生物素 - 亲和素 )

C. High-throughput detection of viral nucleic acids- DNA chips(DNA 芯片 )

B.PCR Technology HCV, HIV, CMV, and HPV

Annealing ( 低温退火 )

Extension ( 适温延伸 )

Denaturation ( 高温变性 )

Page 11: Diagnosis, prevention and treatment Xiao-Kui Guo PhD

Rapid diagnosis of viral infection A. Morphologic observation

1. Optical microscope 2. Electron microscopy (EM) 3.Immune electron microscopy (IEM) ( 免疫电镜技

术 )

B. Immunological detection method 1. Virus antigen detection 2. Measurement of virus specific IgM antibody 3. Multiple viral antigen/antibody detection assay 4. PCR

Page 12: Diagnosis, prevention and treatment Xiao-Kui Guo PhD

Section B

The prevention of the viral infection

Page 13: Diagnosis, prevention and treatment Xiao-Kui Guo PhD

Prevention of viral infection(病毒感染的免疫预防)

Immune prevention is a method of artificial inoculation of biological agents to achieve the aim of disease prevention, control and eradication of infectious diseases in human.

"Artificial Immunity"

Page 14: Diagnosis, prevention and treatment Xiao-Kui Guo PhD

Artificial active immunization

A. Live attenuated vaccines ( 减毒活疫苗 ) B. Inactivated vaccine C. Genetically engineering vaccine D. Nucleic acid vaccine ( 核酸疫苗 )

Page 15: Diagnosis, prevention and treatment Xiao-Kui Guo PhD

Comparison of characteristics of killed and live viral vaccines.

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Artificial passive immunization

Artificial passive immunization ( 人工被动免疫 ) is defined as host is directly injected with passive immunization artificial biological agents.

A. "Normal" immune globulin ( 普通免疫球蛋白 ) B. Antivirus serum C. Immunomodulating agents :the transfer factor, interleukin

( 白细胞介素 ), thymosin ( 胸腺素 ), interferon ( 干扰素 ) etc.

Page 18: Diagnosis, prevention and treatment Xiao-Kui Guo PhD

Section C

Anti-virus treatment

Page 19: Diagnosis, prevention and treatment Xiao-Kui Guo PhD

Antivirus therapy (抗病毒治疗 )

Large-scale vaccination programs have represented the most effective antiviral defense to some viruses, but to many other virus still lack of specific vaccines. And vaccines provide modest to no therapeutic effect in individuals that are already infected.

Antiviral therapy includes two combined strategies: Agents are used to inhibit viral replication directly; Using drugs to stimulate host immune responds so

that the infected cells could be killed.

Page 20: Diagnosis, prevention and treatment Xiao-Kui Guo PhD

Chemotherapy ( 化学治疗 )

A . Inhibition of Viral Entry, uncoating and release

B. Inhibition of viral replication and transcription 1.Nucleoside analogs ( 核苷类类似物 )

(1) Acyclovir (2) 3'-azido-deoxythymidine (3) Lamivudine (4) Ribavarin (5) Idoxuridine (6) Adenine arabinaside

2. Nonnucleoside RT Inhibitors C. Inhibition of viral protein modification

Page 21: Diagnosis, prevention and treatment Xiao-Kui Guo PhD

Immunotherapy

A. Immunomodulator 1. Immunomodulating agents 2. Toll-like receptor 3. Therapeutic vaccine

B. Modulation and retransfusion of immunocyte ( 免疫细胞的修饰与回输 )

Page 22: Diagnosis, prevention and treatment Xiao-Kui Guo PhD

Gene therapy

Gene therapy ( 基因治疗 ) is the introduction of genetic material into cells for therapeutic purposes.

Many human diseases are caused by the absence or inappropriate presence of a protein.

Technology: gene identification, gene expression and gene delivery

Page 23: Diagnosis, prevention and treatment Xiao-Kui Guo PhD

The end

Thanks