Practical Virology Lab. (1)

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Presentation transcript:

Practical Virology Lab. (1) Introduction

What is the virus ?? Particle composed of inner core of either DNA or RNA surrounded by a protein coat Must replicate within the cell Replicate in a different manner from cells (not binary fission)

Methods of Diagnosing Viral Infections 1. Virus isolation 2. Direct detection of the virus a. Detection of the virus Ag (antigenaemia assay, ELISA, latex agglutination) b. Immune EM c. Detection of the virus nucleic acid (PCR, In situ hybridization) d. Immunocytochemistry, and immunohistochemistry 3. Indirect (Serology) … IgM, IgG (titer)

Surface protein of the virus: Antigen mounts an immune response Determinant of type specificity (eg. Polio virus type 1, 2, and 3) Serological diagnosis (detection of the antibody against this antigen)

Virus Isolation Using three living systems

Isolation of the virus using three living systems: 2. Embryonated egg Tissue Culture 3. Lab. Animals

Why we isolate the virus in a living system ?? Viruses must replicate within the cells because they can not generate energy or synthesize protein

Types of tissue culture

1. Primary tissue culture: from adult human or animal eg 1. Primary tissue culture: from adult human or animal eg. (monkey kidney), can be sub-cultured once or twice. Advantages -Normal cells -Contain the original number of chromosomes Disadvantages -Can be sub-cultured once or twice -Should be prepared freshly each time -Requires continuous organ supply

Advantages Disadvantages 2. Semi-continuous: from human fetal tissue, eg. human embryo lung fibroblasts, MRC-5, W138, can be sub-cultured 20-50X. Advantages 20-50 passages Susceptibility for many viruses Can grow fastidious viruses Disadvantages Difficult to be obtained

3. Continuous (Cell line): from human or animal tumors, eg 3. Continuous (Cell line): from human or animal tumors, eg. Vero, HEp-2, HeLa, Hepatoma cell line, can be sub-cultured indefinitely. Advantages Can be sub-cultured indefinitely Easy to obtain Disadvantages Malignant cells (could express altered phenotype, different number of chromosomes)

SARS- infected Vero cells

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