diagnosis, prevention and treatment
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Diagnosis, prevention and treatment. Xiao-Kui Guo PhD. Content. Lab diagnosis of the viral infection Prevention of the viral infection Anti-virus treatment. Section A Lab Diagnosis of the viral infection. Methods of the diagnosis. - PowerPoint PPT PresentationTRANSCRIPT
Diagnosis, prevention and treatment
Xiao-Kui Guo PhD
Content
Lab diagnosis of the viral infection Prevention of the viral infection Anti-virus treatment
Section A
Lab Diagnosis of the viral infection
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)
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
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
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
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)
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 )
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 )
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
Section B
The prevention of the viral infection
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"
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 )
Comparison of characteristics of killed and live viral vaccines.
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.
Section C
Anti-virus treatment
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.
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
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 )
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
The end
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