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Questions and Answers
Questions and Answers
Which of the following is NOT a characteristic of viruses?
Which of the following is NOT a characteristic of viruses?
- Possess a protein coat
- Multiply inside living cells
- Contain either DNA or RNA
- Undergo binary fission (correct)
Viruses are sensitive to antibiotics.
Viruses are sensitive to antibiotics.
False (B)
Which of the following best describes a virion?
Which of the following best describes a virion?
- A virus that infects and kills bacteria
- The protein shell that surrounds and protects the viral genome
- The complete, infectious virus particle (correct)
- The structural protein units that make up the capsid
The proteinaceous subunit that makes up that make up the viral capsid is called a __________.
The proteinaceous subunit that makes up that make up the viral capsid is called a __________.
What is the function of the viral capsid?
What is the function of the viral capsid?
All viruses contain a lipoprotein envelope.
All viruses contain a lipoprotein envelope.
In the context of viruses, what is a nucleocapsid?
In the context of viruses, what is a nucleocapsid?
Match the viral structure with its description.
Match the viral structure with its description.
Which of the following describes viruses that do NOT have an envelope?
Which of the following describes viruses that do NOT have an envelope?
Naked viruses are more stable in hostile environments than enveloped viruses.
Naked viruses are more stable in hostile environments than enveloped viruses.
How do enveloped viruses typically acquire their envelope?
How do enveloped viruses typically acquire their envelope?
Which of the following is a mode of transmission more commonly associated with enveloped viruses?
Which of the following is a mode of transmission more commonly associated with enveloped viruses?
Viruses that infect bacteria are known as __________.
Viruses that infect bacteria are known as __________.
What is a key difference in the genome structure between retroviruses and other viruses?
What is a key difference in the genome structure between retroviruses and other viruses?
A virus can have both DNA and RNA within its genome.
A virus can have both DNA and RNA within its genome.
Which of the following is NOT a possible symmetry of a nucleocapsid?
Which of the following is NOT a possible symmetry of a nucleocapsid?
What process is required for retroviruses to integrate their RNA genome into the host cell's DNA?
What process is required for retroviruses to integrate their RNA genome into the host cell's DNA?
What is the term for viruses' process by which they exit the cell
What is the term for viruses' process by which they exit the cell
Viral surface proteins mediate attachment to host cell __________, determining the host and organ specificity of the virus.
Viral surface proteins mediate attachment to host cell __________, determining the host and organ specificity of the virus.
Which phenotypic characteristic is NOT typically used to classify viruses?
Which phenotypic characteristic is NOT typically used to classify viruses?
RNA viruses generally have higher mutation rates than DNA viruses.
RNA viruses generally have higher mutation rates than DNA viruses.
Match Baltimore's seven classes of virus with properties of their nucleic acid
Match Baltimore's seven classes of virus with properties of their nucleic acid
Which of the following is an example of vertical transmission of a virus?
Which of the following is an example of vertical transmission of a virus?
Give an example of a virus that causes infection via an animal bite.
Give an example of a virus that causes infection via an animal bite.
What is 'cell tropism' in the context of viral infections?
What is 'cell tropism' in the context of viral infections?
The time from exposure to a virus until the onset of clinical disease is known as the __________ period.
The time from exposure to a virus until the onset of clinical disease is known as the __________ period.
What type of immune cells are primarily responsible for clearing viral infections?
What type of immune cells are primarily responsible for clearing viral infections?
Antibodies play a major role in recovery from an active viral infection.
Antibodies play a major role in recovery from an active viral infection.
What is one of the first non-specific host defenses triggered by viral infection?
What is one of the first non-specific host defenses triggered by viral infection?
How do natural killer (NK) cells contribute to viral defense?
How do natural killer (NK) cells contribute to viral defense?
In the context of viral diagnosis, what is the purpose of electron microscopy?
In the context of viral diagnosis, what is the purpose of electron microscopy?
In PCR, __________ are used to amplify fragments of viral nucleic acid from samples to be tested.
In PCR, __________ are used to amplify fragments of viral nucleic acid from samples to be tested.
Which viral diagnostic tests depends on a fluorescein-labeled antibody staining viral antigens, so that cells fluoresce under UV illumination?
Which viral diagnostic tests depends on a fluorescein-labeled antibody staining viral antigens, so that cells fluoresce under UV illumination?
Which detection method would you chose for a current viral infection?
Which detection method would you chose for a current viral infection?
Cell cultures are needed for the growth of viral cultures.
Cell cultures are needed for the growth of viral cultures.
Which of the following human viruses is not dsDNA virus?
Which of the following human viruses is not dsDNA virus?
Poxviridae are known to include Herpes simplex viruses types 1 and 2, varicella-zoster virus, Epstein-Barr virus, and cytomegalovirus.
Poxviridae are known to include Herpes simplex viruses types 1 and 2, varicella-zoster virus, Epstein-Barr virus, and cytomegalovirus.
Why is herpes simplex often latent?
Why is herpes simplex often latent?
List 3 symptoms of herpes type 1
List 3 symptoms of herpes type 1
Which cell type do leukocyte sites not include for latency purposes?
Which cell type do leukocyte sites not include for latency purposes?
Match these key virus infections to one of their key characteristics
Match these key virus infections to one of their key characteristics
The measles vaccine has not reduced cases of Chickenpox or Fifth disease
The measles vaccine has not reduced cases of Chickenpox or Fifth disease
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Flashcards
Flashcards
Virion
Virion
The complete infectious virus particle, consisting of a nucleic acid genome (DNA or RNA) surrounded by a protein coat.
Capsid
Capsid
A protein shell that surrounds and protects the viral genome, made up of multiple identical protein subunits called capsomeres.
Capsomeres
Capsomeres
The structural protein units that make up the capsid.
Nucleocapsid
Nucleocapsid
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Envelope (Viral)
Envelope (Viral)
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Glycoproteins
Glycoproteins
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Bacteriophage/Phage
Bacteriophage/Phage
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Viral Core
Viral Core
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Viral Genome
Viral Genome
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Obligate intracellular parasite
Obligate intracellular parasite
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Cytopathic Effect (CPE)
Cytopathic Effect (CPE)
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Naked Viruses
Naked Viruses
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Enveloped Viruses
Enveloped Viruses
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Viral Surface Proteins
Viral Surface Proteins
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Matrix Proteins
Matrix Proteins
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Internal Proteins (Viral)
Internal Proteins (Viral)
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Attachment and Entry (Viral Replication)
Attachment and Entry (Viral Replication)
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Uncoating (Viral Replication)
Uncoating (Viral Replication)
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Transcription (Viral Replication)
Transcription (Viral Replication)
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Genome Replication (Viral)
Genome Replication (Viral)
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Translation (Viral Replication)
Translation (Viral Replication)
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Virion Assembly (Viral)
Virion Assembly (Viral)
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Virion Release (Viral)
Virion Release (Viral)
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Vertical Transmission
Vertical Transmission
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Horizontal Transmission
Horizontal Transmission
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Cell Tropism
Cell Tropism
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Incubation Period
Incubation Period
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Non-specific Defense (Viral)
Non-specific Defense (Viral)
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Natural Killer (NK) Cells
Natural Killer (NK) Cells
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Interferons
Interferons
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Specific Host Defenses (Viral)
Specific Host Defenses (Viral)
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Antiviral Antibody
Antiviral Antibody
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Cytotoxic T-lymphocytes
Cytotoxic T-lymphocytes
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Electron Microscopy (Viral Diagnosis)
Electron Microscopy (Viral Diagnosis)
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Immuno-electron microscopy
Immuno-electron microscopy
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PCR (Polymerase Chain Reaction)
PCR (Polymerase Chain Reaction)
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Immunofluorescence (IF) methods
Immunofluorescence (IF) methods
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ELISA (Enzyme-Linked Immunosorbent Assay)
ELISA (Enzyme-Linked Immunosorbent Assay)
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Virus Identification in Cell Culture
Virus Identification in Cell Culture
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Hemagglutination Test
Hemagglutination Test
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Study Notes
Study Notes
Module Contents
- Module of Microbiology II includes Medical Bacteriology and Medical Virology.
Medical Virology Introduction
- Aims to give students virology knowledge, understand genesis of diseases, and know diagnosis and prevention.
- Covers virus classification, isolation, culture, pathogenicity, and serological reaction mechanisms.
Course Outline
- Includes the history, terminology, structure, characteristics, classification, and transmission of viruses.
- Covers virus replication and pathogenesis, host defense, viral infections (CNS, GIT, RT, STIs, Hepatitis, Hemorrhagic fever), and diagnosis.
What is a Virus?
- Submicroscopic entity with a single nucleic acid surrounded by a protein coat that replicates inside living cells of bacteria, animals, humans, or plants.
- In short, a virus is an obligate intracellular organism.
Viruses
- Contain a single type of nucleic acid, either DNA or RNA.
- Have a protein coat (sometimes enclosed by an envelope of lipids, proteins, and carbohydrates) around the nucleic acid.
- Multiply inside living cells using the cell's synthesizing machinery.
- Cause synthesis of specialized structures to transfer viral nucleic acid to other cells.
Terminology
- Virion: Complete, infectious virus particle, including a genome comprising one or a few molecules of DNA or RNA.
- Capsid: Protein shell surrounding and protecting the genome built of multiple protein subunits called capsomeres.
- Capsomeres: Structural protein units making up the capsid, capsids being either icosahedral or tubular in shape.
- Nucleocapsid: Capsid and the enclosed nucleic acid, together.
- Envelope: Lipoprotein membrane that comes from the host cell's plasma membrane, surrounds some viruses.
- Glycoproteins: Proteins in the virion's envelope, typically glycosylated.
- Bacteriophage/phage: Virus that infects and kills bacteria without harming human and animal cells (e.g., Escherichia coli).
General Characteristics of a Virus
- Filterable agents, obligate intracellular parasites.
- Viral genome is RNA or DNA, but not both.
- May or may not have a lipoprotein envelope.
- Metabolically inert and can only replicate inside a host cell.
- Very small, sizes range between 20 - 200 nm, beyond the resolving power of the light microscope.
Comparisons of Viruses & Bacteria
- Bacteria grow on artificial media, viruses never grow on artificial media.
- Bacteria divide by binary fission, viruses never divide by binary fission.
- Bacteria contain DNA & RNA, Virus either contains DNA or RNA.
- Bacteria contain synthesis machinery, viruses dont
- Bacteria are sensitive to antibiotics, viruses aren't
Structucture Of Viruses
- Viruses possess either:
- DNA and RNA
- Plasma Membrane
- Binary Fission
- Pass through bacteriological filters
- ATP-Generating Metabolism
- Ribosomes
- Sensitive to Antibiotics
- Sensitive to Interferon
Viral Core
- Viral Core us the viral nucleic acid genome, found in the center of the virion.
- It control the viral heredity and variation
- Responsible for the infectivity
Genome
- The genome of a virus can be either DNA or RNA.
- DNA can be double stranded (ds) or single stranded (ss).
- RNA can be segmented or non-segmented single stranded (ss) or the double stranded (ds) is only Linear (only Reovirus family)
- All viruses have one copy of their genome (haploid) except retroviruses, which are diploid.
Viral Capsid
- The protein shell or coat, that encloses the nucleic acid genome where it :
- Protects the viral nucleic acid
- Participate in the viral infection
- Share the antigenicity
Nucleocapsid
- The core of a virus particle consisting of the genome plus a complex of proteins:
- Complex of proteins = Structural proteins + Non-Structural proteins (Enzymes & Nucleic acid binding proteins)
Symmetry of Nucleocapsid
- Helical/spiral
- Cubic/Icosahedral
- Complex
Envelope
- The viral envelope consists of a membrane that contains lipid derived from the host cell and proteins encoded by the virus.
- The envelope is acquired as the virus exits from the cell in a process called budding.
- Viruses-encoded glycoproteins are exposed on the surface of the envelope.
Functions of Envelope
- Antigenicity, Infectivity and Resistance
- Not all viruses have the envelope and are divided into:
- Enveloped virus
- Naked virus
Properties of Naked Viruses
- Stable in hostile environments and not damaged by drying, acid, detergent, and heat.
- Released by lysis of host cells and can infect the GI tract and survive the acid and bile.
- Spread easily via hands, dust, fomites and can retain infectivity when dry .
- Needs neutralizing of mucosal and systemic antibodies to control infection.
- Survive longer in the environment and are transmitted by indirect means, like the fecal-oral route.
Properties of Enveloped Viruses
- Labile in dry, arid environments and damaged by drying, acid, detergent, and heat.
- Pick up new cell membrane during multiplication and inserts new virus-specific proteins after assembly.
- Released by budding and are transmitted by direct contact via blood and body fluids.
Viral Proteins
- Viral surface proteins mediate attachment to host cell receptors, determining host and organ specificity.
- They are targets of antibodies that prevent virus attachment and neutralize viral replication.
- Matrix proteins mediate interaction between viral nucleocapsid and envelope proteins.
- Viruses have internal proteins, including DNA or RNA polymerases.
Classification of Viruses
- Mainly classified by phenotypic characteristics like morphology, nucleic acid type, replication mode, host organisms, and disease type.
- Classified according to their genome and structures.
- This includes:
- Nucleic acid type (RNA or DNA, single or double stranded, segmented or non-segmented, linear or circular).
- Virion Structure (Symmetry icosahedral, helical, or complex) with or without envelope.
- Replication strategy.
Viruses are divided into two large groups:
- RNA containing viruses.
- DNA containing viruses.
Classification of Viruses based on
- Morphology, Chemical composition, and mode of replication with:
- Morphology: Focuses on the size and shape, and structure of the genome(eg: helical symmetry, icosahedral symmetry, etc).
- Chemical composition and mode of replication: focus on the genome of a virus (either DNA or RNA, which may be single stranded (ss) or double stranded (ds), linear or circular). Error rate and mutation rates.
Groups of Viruses
- Examples of groups include:
- ds DNA VIRUSES: Parpovaviruses, Adenoviruses, Herpes viruses, Poxviruses,, and Iridoviruses.
- ss DNA VIRUSES: Parvoviruses
- (+) SENSE SSRNA VIRUSES : Picornaviruses, togaviruses, coronaviruses, caliciviruses, and hepatitis E viruses (-)VE SENSE SSRNA VIRUSES : Orthomyxoviruses, paramyxoviruses, bunyaviruses, arenaviruses, and rhabdo-viruses,
- ds RNA VIRUSES : Reoviridiae
- Retroviruses
Classification of Viruses: ICTV
- International Committee on Taxonomy of Viruses (ICTV) classifies viruses into 3 orders, 73 families, 287 genera, and over 2000 species.
Baltimore Classification
- Created by David Baltimore, based on method of viral m-RNA synthesis:
- 1- ds-DNA viruses
- 2- ss-DNA viruses
- 3- ds- RNA viruses
- 4- ss-RNA viruses with positive strands ( positive polarity).
- 5- ss-RNA viruses with negative strands(negative polarity).
- 6- ss-RNA viruses associated with the enzyme reverse transcriptase (Retroviruses)
- 7-ds-DNA-RT viruses (ex;Hepadnaviruses)
Examples of Baltimore Groups
- Double stranded DNA families = Poxviridae (smallpox), Herpesviridae (Herpes simplex virus type 1), Hepadnaviridae (Hepatitis B), Adenoviridae (Adenovirus 40 & 41), Papovaviridae.
- Single stranded DNA families = Parvoviridae
- Double stranded RNA families = Reoviridae ex; Rota virus;
- Single stranded RNA families with positive strands = Picornaviridae ex; Polio virus, Caliciviridae ex; Norwalk virus, Astroviridae ex; Astrovirus,, Coronaviridae ex; Coronavirus, Flaviviradae ex ;Hepatitis C virus, yellow fever virus, Togaviridae ex; Rubella virus
- Single stranded RNA families with negative strands = Orthomyxoviridae ex; Infuenzaviruses, Paramyxoviridae ex; Parainfluenza, Rhabdoviridae ex; Rabies virus,Filoviridae. Ex; Ebola virus
- Single stranded RNA viruses associated with the enzyme reverse transcriptase = Retroviruses ex; HIV
Transmission of Viruses
- Mother to offspring = Vertical transmission
- All other transmission modes (fecal-oral, respiratory aerosol, insect bite) = Horizontal transmission.
- Can be from human to human or from animal to human.
- Examples of transmission Include:
- Respiratory transmission ex; Influenza A virus
- Faecal-oral transmission ex; Enterovirus
- Blood-borne transmission ex; Hepatitis B virus
- Sexual Transmission ex; HIV
- Animal bite ex; Rabies virus
- Insect vectors ex: Yellow fever
Viral Replication Steps
- Important replication steps comprise of:
- Attachment and entry: Interaction of virion structures with host cell receptors defines the host species. This includes specific receptor on the cell surface.
- Uncoating: Protein coat of the virion dissociates
- Transcription: Virus mRNA is produced using either cellular or virus-coded enzymes.
- Genome replication occurs in the cytoplasm or nucleus. Enzymes may be encoded by the virus or the host cell.
- Translation is where cellular machinery synthesises various viral proteins using ribosomes.
- Virion Assembly: Newly formed virus proteins and nucleic acid assemble to produce new virus particles.
- Virion release which is Enveloped viruses exit by budding, picking up the envelope and for Naked viruses it results in lysis of the cell. Whereby RNA viruses replicate in cytoplasm except HIV, Influenza and DNA viruses replicate in the Nucleus except poxvirus
How Viruses Cause Diseases
- Viruses infect all types of living organisms, from bacteria to humans, requiring the right cell receptor.
- Enter the body by inhalation, ingestion, sexually, parenteral or inoculation, and vertical transmission
- Infection can either remain localized to the site of entry, or it may cause a disseminated infection according to the site of target.
- The virus initially replicates at the site of entry, then enters the blood (viraemia) or lymphatics, other viruses may travel up the axon to infect the central nervous system
- There is a time from exposure to the onset of clinical disease called the incubation period: Viruses that cause localized infections have short incubation periods (less than 7 day), while in disseminated infections, incubation is longer
- The immune response required is the action of specific cytotoxic T lymphocytes (T8). Virus-specific antibody levels rise during the course of the infection Specific antibodies play a very important role in preventing reinfection of the host with the same virus
- There is the potential for certain viruses to evade the immune response and establish persistent infections in their host
Viral Pathogenesis mechanisms
- Implantation of virus at the portal of entry
- Local replication.
- Spread to target and shedding of virus into the environment.
Viral Pathogenesis Effects
- To produce disease, viruses must enter a host, come in contact with susceptible cells, replicate, and produce cell injury
- There are four main effects of virus infection on the cell:
- No apparent morphological or functional change.
- Fusion of cells to form multinucleated cells.
- Malignant transformation.
- Death .
General Viral Pathogenesis
- Cell destruction following virus infection which leads to death of the infected cells or the cytopathic effect (CPE)
- Virus-induced changes to cellular gene expression can cause virus-induced tumours. in the host cell . A hallmark of viral infection of the cell is the cytopathic effect
- Immuno-pathogenesis is where symptoms of viral diseases are caused by the immune system rather than the virus this includes the killing of virus-infected cell by the attack of cytotoxic T cells or the form virus-antibody complexes that are deposited in tissues.
Host Defenses Against Viral Infection
- Number of host defences contribute to the prevention and/or elimination of viral infections and fall into two major categories:
- non specific defense: include anatomical barriers, mucus membrane and cilia clearance, after infection factors such as fever and inflammatory act to reduce viral replication. The most important are interferons and natural killer cells
- Specific host defences: Natural Killer cells kill virus-infected cells by secreting perforins and granzymes, which cause apoptosis and interferons are released by viral infected cell and inhibit the growth of viruses by blocking the translation of viral proteins
Host Defenses Specific
- Include antiviral antibody, which may prevent adsorption to target cells and prevent reinfection of the host with the same virus
- Cytotoxic T-lymphocytes, which recognize virally-infected cells and destroy them, reducing viral production.
Viral Diagnosis Approaches
- Observation of virus particles by electron and immunoelectron microscopy
- Detection of viral nucleic acids in blood or patient's cells
- Detection of viral antigens in blood or body fluids
- Detection of virus-specific antibodies in the blood
- Identification of the virus in cell culture
Electron Microscopy for Viral Diagnosis
- Used for rapid identification of morphologically distinctive virions, with immuno-electron microscopy enhancing the test using specific immune serum.
- Useful where virus cannot be cultivated.
Detection of Viral Nucleic Acid
- In PCR, specific oligonucleotides (primers) are used to amplify fragments of viral nucleic acid from samples.
- PCR involves denaturation, annealing, and extension steps using a thermo-stable DNA polymerase.
- Viral RNA can also be used, by including a reverse transcription (RT)-PCR step in which it’s converted into DNA.
Detection of Viral-Specific Antibodies
- Immunofluorescence methods are widely used for rapid virus infection diagnosis
- This includes detecting of viral antigens in clinical specimens, as well as the detecting of virus-specific IgG, IgA, or IgM antibody,
- There, using of fluorescencein- labeled antibody stains, specimens containing specific virus antigens, allowing cell fluoresces. under UV
Identification of the Virus in Cell Culture
- The growth of viruses requires cell cultures, because viruses replicate only in living cells:
- Methods for growing virus includes:
- Animals, Embryonated eggs & Monolayer cell
- Transport or storage at 4°C is acceptable.
- Animals, Embryonated eggs & Monolayer cell
- Virus in cell culture produces a cytopathic effect (CPE).
CPE is a change in appearance of the cells in size, shape, forming multinucleated giant cells and Inclusion bodies. These are observed with light microscopy and can be related to:
- Total destruction of host cell monolayer ex: enteroviruses
- Subtotal destruction ex; some paramyxoviruses
- Focal Degeneration ex; herpes virus and poxvirus;Swelling and clumping ex; Adenoviruses
- Cytoplasmic Vacuoles ex; Paramyxovirus
- Cell fusion (Syncytia) ex; Herpes viruses, paramyxoviruse
- There is the used of known antibody to identify the virus is achieved by :
- Example hemagglutination test which are limited to Attachment of erythrocytes to the cell of viruses with a hemagglutinin. protein on their envelope, such as mumps, parainfluenza, and influenza viruses and use of Hemagglutination inhibition test
- If the virus and antibody are similar, the virus is blocked from attaching to the erythrocytes and it means the test in positive for the specific antibody being tested this means the viruses agglutinate red.blood cell. - There is the use of Fluorescent-Antibody Assay of viral infected cell the Florescence signal is seen in virus infected cells .and use. of ELISA to determine viral amounts - In brief the presence of IgM is used to diagnose current infection and IgG is used to determine immunity
Disinfection and Inactivation of Viruses
- Heat can be used at 56°C for 30 minutes or at 100°C for just a few seconds
- Drying and Ultra-violet irradiation can be used for enveloped viruses by not for non-enveloped viruses
- lipid solvents (Chloroform, Ether, Alcohol and Oxidizing and reducing agents
- Phenols do kill viruses via inactivation
Medically Important DNA Viruses
- Families of DNA viruses include:
- Poxviridae ex; Variola virus,.
- Herpesviridae, Epstein-Barr virus, cytomegalovirus,
- Adenoviridae; Adenovirus, Hepadnaviridae; Hepapillomavirus, Polyomaviridae;
- Parvoviridae; B19
Herpesviruses
- Herpesviruses are icosahedral shaped that are 90 to 100 nm
- Members of the Herpesviridae family.
- Except for neonates, those infections are more severe in adults than in children
- Latent infections in Site of Leukocytes and peripheral nerves.
- Reactivation is triggered resulting in is milder than primary infection ex shingles
- Where herpesvirus includes VZV, HSV-1 and -2, CMV
Examples of herpesvirus infections
- Herpes simplex viruses are Gingivostomatitis, recurrent oral-labial and cornea infection
- Herpes simplex virus type 2 the causes with Genital herpes
- Varicella zoster cause chickenpox that can Reactivation as shingles where Epstein-Barr virus cause Hetero-positive mononucleosis in B lymphocytes. Cytomegalovirus that gives fever. and Human herpesvirus 6 and 7cause Roseola that is found Peripheral cells
- Human herpesvirus 8, causes Kaposi’s sacroma found Peripheral cells
Herpes simplex virus type 1 and 2
- HSV-1 causes mouth lesions and fever blisters.
- Most cases are mild, with mild fever and general malaise sometimes
- Infections can be asymptomatic that is diagnosed using :Symptoms
- Diagnostic testing use Immunologic assays HSV can grow in continuous cell lines e.g., HEp-2 and A549 and established cell lines e.g., MRC-5
- HSV-2 is a causative agent of genital herpes and can transferred from mother at birth and can cause Eye Infections and cause damage the nervous system which in diagnosed in immunologic assays that detect viral antigens
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