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Medical Virology
Dept. of Medical Microbiology and Parasitology
Shanghai Second Medical University
Shanghai, China.
E.mail: ( [email protected] )
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General Virology
Conception Viruses Virion
Size and ShapeStructure
Replication Viral Variation Classification
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Conception Virology is the bioscience for study of
viral nature,and the relationship between viruses and hosts. Viruses often cause serious diseases, relate to some cancers and congenital deformities, also can be used as tool for genetic engineering.
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3000BC
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History Smallpox was endemic in
China by 1000BC. In response, the practice of variolation was developed. Recognizing that survivors of smallpox outbreaks were protected from subsequent infection, variolation involved inhalation of the dried crusts from smallpox lesions like snuff, or in later modifications, inoculation of the pus from a lesion into a scratch on the forearm of a child.
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Definition of Virus Viruses may be defined as acellular
organisms whose genomes consist of nucleic acid, and which obligately replicate inside host cells using host metabolic machinery and ribosomes to form a pool of components which assemble into particles called VIRIONS, which serve to protect the genome and to transfer it to other cells
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Viral Properties
Viruses are inert (nucleoprotein ) filterable Agents
Viruses are obligate intracellular parasites Viruses cannot make energy or proteins
independent of a host cell Viral genome are RNA or DNA but not both. Viruses have a naked capsid or envelope with
attached proteins Viruses do not have the genetic capability to
multiply by division. Viruses are non-living entities
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Consequences of Viral Properties
Viruses are not living Viruses must be infectious to endure in nature Viruses must be able to use host cell processes
to produce their components (viral messenger RNA, protein, and identical copies of the genome)
Viruses must encode any required processes not provided by the cell
Viral components must self-assemble
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Challenges the way we define life
viruses do not respire, nor do they display irritability 应急性 ; they do not move they do not grow they do most certainly reproduce, and may
adapt to new hosts.
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Size and Shape
Methods Size of Viruses Shapes of Viruses
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Methods of Analysis
Electron microscopy : The resolution is 5nm (1nm = 10-9 m) X-ray crystallography
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Size of Viruses
A small virus has a diameter of about 20nm.
Parvovirus
A large virus have a diameter of up to 400nm.
Poxviruses
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Shape of Viruses
Spherical Rod-shaped Brick-shaped Tadpole-shaped Bullet-shaped Filament
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Shapes of Viruses:Spherical
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Shapes of Viruses :Rod-shaped
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Shapes of Viruses :Brick-shaped .
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Tadpole-shaped
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Shapes of Viruses :Bullet-shaped
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Shapes of Viruses :Filament
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Structure of Viruses
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Virion the complete infectious
unit of virus particle Structurally mature,
extracellular virus particles.
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Virion
Capsid
Viral core
envelope
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Viral core
Viral core
The viral nucleic acid genome, In the center of the virion, : Control the viral heredity and variation, responsible for the infectivity.
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Genome The genome of a virus can be either DNA or
RNA
DNA-double stranded (ds): linear or circular Single stranded (ss) : linear or
circular RNA- ss:segmented or non-segmented ss:polarity+(sense) or polarity –
(non-sense) ds: linear (only reovirus family)
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DNA RNA
double-stranded
single-stranded
double-stranded
single-stranded
linear
circularlinear
circular linear linear (circular)
single
single
multiple
single
single
multiple
single
multiple
(+)sense (-)sense
single
multiple
single
multiple
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Viral Capsid The protein shell, or coat, that
encloses the nucleic acid genome. Functions: a. Protect the viral
nucleic acid. b. Participate in the viral infection. c. Share the antigenicity
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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)
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Symmetry of Nucleocapsid
Helical Cubic
/Icosahedral Complex
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Helical symmetry
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Helical symmetry
How to assemble
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Helical California Encephalitis Virus
CoronavirusHantavirusInfluenza Virus (Flu Virus)Measles Virus ( Rubeola)Mumps VirusParainfluenza VirusRabies VirusRespiratory Syncytial Virus(RSV)
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Cubic or icosahedral symmetry
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Icosahedral Adeno-associated Virus
(AAV)AdenovirusB19Coxsackievirus - ACoxsackievirus - BCytomegalovirus (CMV)Eastern Equine Encephalitis Virus (EEEV)EchovirusEpstein-Barr Virus (EBV)Hepatitis A Virus (HAV)Hepatitis B Virus (HBV)Hepatitis C Virus (HCV)Hepatitis Delta Virus (HDV)Hepatitis E Virus (HEV)
Herpes Simplex Virus 1 (HHV1)Herpes Simplex Virus 2 (HHV2)Human Immunodeficiency Virus (HIV)Human T-lymphotrophic Virus (HTLV)Norwalk VirusPapilloma Virus (HPV)Polio virusRhinovirusRubella VirusSaint Louis Encephalitis VirusVaricella-Zoster Virus (HHV3)Western Equine Encephalitis Virus (WEEV)Yellow Fever Virus
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Complex Virus Structures
A well known example is the tailed bacteriophages such as T4.
The head of these viruses is cubic with a triangulation number of 7. This is attached by a collar to a contractile tail with helical symmetry.
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T4 Bacteriophage
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Properties of naked viruses Stable in hostile environment Not damaged by drying, acid, detergent, and heat Released by lysis of host cells Can sustain in dry environment Can infect the GI tract and survive the acid and
bile Can spread easily via hands, dust, fomites, etc Can stay dry and still retain infectivity Neutralizing mucosal and systemic antibodies are
needed to control the establishment of infection
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Naked viruses( Non Enveloped )
Adeno-associated Virus (AAV)AdenovirusB19Coxsackievirus - ACoxsackievirus - BEchovirusHepatitis A Virus (HAV)Hepatitis E Virus (HEV)Norwalk Virus
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Envelope A lipid-containing membrane
that surrounds some viral particles.
It is acquired during viral maturation by a budding process through a cellular membrane, Viruses-encoded glycoproteins are exposed on the surface of the envelope.
Not all viruses have the envelope, and viruses can be divided into 2 kinds: enveloped virus and naked virus.
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Functions of envelope
Antigenicity
some viruses possess neuraminidase
Infectivity
Resistance
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Envelope
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Properties of enveloped viruses
Labile in dry , arid environment Damaged by drying, acid, detergent,
and heat Pick up new cell membrane during
multiplication Insert new virus-specific proteins
after assembly Virus is released by budding
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Consequences of Properties for enveloped viruses
Must stay moist Must not infect the GI tract for survival Must be transmitted in the protective,
droplets, secretions, blood and body fluids Must reinfect another host cell to sustain Humoral and cell-mediated immunity are
needed to control the infection
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Enveloped California Encephalitis Virus Coronavirus Cytomegalovirus (CMV) Eastern Equine Encephalitis
Virus (EEEV) Epstein-Barr Virus (EBV) Hantavirus
Hepatitis B Virus (HBV)Hepatitis C Virus (HCV)Hepatitis Delta Virus (HDV)Herpes Simplex Virus 1 (HHV1)
Rotavirus Rubella Virus Saint Louis Encephalitis Virus
Smallpox Virus (Variola)Vaccinia Virus
Herpes Simplex Virus 2 (HHV2)Human Immunodeficiency Virus (HIV)Human T-lymphotrophic Virus (HTLV)Influenza Virus (Flu Virus)Molluscum contagiosumPapilloma Virus (HPV)Polio virusRhinovirusVaricella-Zoster Virus (HHV3)Venezuelan Equine Encephal. Vir. (VEEV)Western Equine Encephalitis Virus (WEEV)Yellow Fever Virus
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Spike or Peplomere
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summarize
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Virion structure
Nucleocapsid(Naked Virus) = DNA or RNA +Structural proteins +Enzymes &Nucleic acid binding proteins
Enveloped Virus =Nucleocapsid+ Viral specific glycoproteins and Host Membrane
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Enveloped
Virus
Naked Virus
Cubic Helical
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CHEMICAL COMPOSITION OF VIRUSES
Viral Protein Viral Nucleic Acid Viral Lipids Viral carbohydrate
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Viral Nucleic Acid
DNA-double stranded (ds): linear or circular
Single stranded (ss) : linear or circular RNA- ss:segmented or non-segmented
ss:polarity+(sense) or polarity –(non-sense)
ds: linear (only reovirus family)
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Viral Protein Structural protein (Capsomere)
Enzyme glycoproteins (spike/viral attachment protein, VAP)
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Culture of Viruses
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System for the propagation of viruses
People Animals : cows;chickens; mice; rats;
suckling mice Embryonated eggs Organ and tissue culture
Organ culture primary tissue culture
cell lines: diploid Tumor or immortalized cell line
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Cytopathic effect, CPE
Inclusion Bodies
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CPE:Viral Cytopathological Effects
Cell death Cell
rounding/Degeneration/AggregationLass of attachments to substrate
Inclusion bodies in the nucleus or cytoplasm, margination of chromatin
Syncytia: multinucleated giant cells caused by virus-induced cell-cell fusion
Cell surface changesViral antigen expression Hemadsorption (hemagglutinin
expression)
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Normal cell and CPE
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Inclusions
Negri body
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TCD50,LD50,ID50
PFU: plaque forming units
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Replication of Viruses
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Replicative cycle As obligate intracellular parasites, Virus
must enter and replicate in living cells in order to “reproduce” themselves. This “growth cycle” involves specific attachment of virus, penetration and uncoating, nucleic acid transcription, protein synthesis, matureation and assembly of the virions and their subsequent release from the cell by budding or lysis
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Initiation Phase
Attachment Penetration Uncoating
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Attachment/Adsorption Virus attaches to the cell surface.
Attachment is via ionic interactions which are temperature-independent.
Viral attachment protein recognizes specific receptors on the cell surface (These may be protein or carbohydrate or lipid components of the cell surface).
Cells without the appropriate receptors are not susceptible to the virus.
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PENETRATION (Virus enters the cell) Virions are either engulfed into vacuoles by
“endocytosis” or the virus envelope fuses with the plasma membrane to facilitate entry
Enveloped viruses Non-enveloped viruses
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Fusing (A) Entry by fusing with
the plasma membrane. Some enveloped viruses fuse directly with the plasma membrane. Thus, the internal components of the virion are immediately delivered to the cytoplasm of the cell.
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HIV
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Endocytosis (B) Entry via endosomes at the cell surface
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influenza virus
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Enveloped viruses Some enveloped viruses require an acid pH for
fusion to occur and are unable to fuse directly with the plasma membrane. These viruses are taken up by invagination of clathrin coated pits into endosomes. As the endosomes become acidified, the latent fusion activity of the virus proteins becomes activated by the fall in pH and the virion membrane fuses with the endosome membrane. This results in delivery of the internal components of the virus to the cytoplasm of the cell
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Non-enveloped viruses Non-enveloped viruses may cross the plasma
membrane directly may be taken up via clathrin-coated pits into
endosomes. They then cross (or destroy) the endosomal membrane.
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un
en
velo
ped
viru
ses
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UNCOATING Nucleic acid has to be sufficiently uncoated that virus
replication can begin at this stage. When the nucleic acid is uncoated, infectious virus particles cannot be recovered from the cell - this is the start of the ECLIPSE phase - which lasts until new infectious virions are made
Uncoating is usually achieved by cellular proteases “opening up” the capsid
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BIOSYNTHESIS
genome synthesis mRNA production protein synthesis
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Flow of events during the replication of Hepadna viruses
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Flow of events during the replication of herpesviruses
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Flow of events during the replication of reoviruses.
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Flow of events during the replication of togaviruses
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Flow of events during the replication of orthomyxoviruses and
paramyxoviruses.
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Flow of events during the replication of retroviruses
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Maturation assembly release
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Maturation The stage of viral replication at which a virus
particle becomes infectious; nucleic acids and capsids are assembled together.
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ASSEMBLY
The stage of replication during which all the structural components come together at one site in the cell and the basic structure of the virus particle is formed.
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RELEASE Disintegration : naked virus cause the host cell
lysis Budding: enveloped viruses
Budding viruses do not necessarily kill the cell. Thus, some budding viruses may be able to set up persistence
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Assembly
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Products of viral replication
Virion DEFECTIVE VIRUS ABORTIVE INFECTION integration
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DEFECTIVE VIRUS deficiency in some aspects of replication, but
interfering the replication of normal viruses ABORTIVE INFECTION When a virus infects a cell (or host), but
cannot complete the full replication cycle ( not biosynthesize their components or not assemble virions.), i.e. a non-productive infection.
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INTERFERENCE Interferon, IFN Defective interfering particle, DIP
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Viral Genetics
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Genome The genome of a virus can be
either DNA or RNA
DNA-double stranded (ds): linear or circular Single stranded (ss) : linear or circular RNA- ss:segmented or non-segmented ss:polarity+(sense) or polarity –(non-
sense) ds: linear (only reovirus family)
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Virus Genomes
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The Structure & Complexity of Virus Genomes
The nucleic acid comprising the genome may be single-stranded or double-stranded, & in a linear, circular or segmented configuration. Single-stranded virus genomes may be:
positive (+)sense, i.e. of the same polarity (nucleotide sequence) as mRNA
negative (-)sense ambisense - a mixture of the two.
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The Structure & Complexity of Virus Genomes
any virus genome will usually include the following:
Composition - DNA or RNA, single-stranded or double-stranded, linear or circular.
Size & number of segments. Terminal structures. Nucleotide sequence. Coding capacity - open reading frames. Regulatory signals - transcription enhancers,
promoters & terminators.
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The Structure & Complexity of Virus Genomes
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Transfection Infection of cells caused by nucleic
acid alone
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Variation
There are two important variation which relate well with medical practices
Antigenicity variation: In most viruse the antigenicity is stable but in some viruses such as influenze virus the antigenicity may vary and cause the disease to epidemic.
Virulence variation(Virulent viruses): Less virulent viruses always used in prevention.
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Mutation
MutantVarianttemperature sensitive(ts) mutant
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Interactionswhen two genetically distinct viruses infect a cell 3 different phenomena can ensue
Recombination /Reassortment Complementation Phenotypic mixing
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Recombination dsDNA viruses Reassortment ( segmented
genomes) RNA viruses: influenza
virus
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Complementation
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Phenotypic mixing The genome of virus A can be
coated with the surface protein of virus type B
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Classification of Viruses
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CLASSIFICATION OF VIRUSES-------basis of classification
Virion morphology Physicochemical properties of the virion Virus genome properties Virus protein proteries Genome organization and replication Antigenic properties Biologic properties
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CLASSIFICATION OF VIRUSES
By 1995 --71 families, 11 subfamilies --164 genera For humans and animals --24 families, --DNA: 7; RNA: 17 for humans
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Survey of DNA-containing Viruses
Parvoviruses: human parvovirus B19
Papovaviruses: papillomaviruses
Adenoviruses: 47 types infect humans
Herpesviruses: human herpesvirus 1-8
Poxviruses: smallpox; vaccinia
Hepadnaviruses: HBV
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Survey of RNA-containing Viruses
Picornaviruses Astroviruses Caliciviruses Reoviruses Arboviruses Togaviruses Flaviviruses Arenaviruses Coronaviruses: SARS
Retroviruses Bunyaviruses Othomyxoviruses Paramyxoviruses: Rhabdoviruses:rabies
virus
Bornaviruses: BDV Filoviruses Other viruses Viroids
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DNA and RNA Viruses 嗜肝病毒科( Headnasviridae ) 逆转录病毒( Retroviridae )
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Viroids( 类病毒 )
Viroids are small (200-400nt), circular RNA molecules with a rod-like secondary structure which possess no capsid or envelope which are associated with certain plant diseases. Their replication strategy like that of viruses - they are obligate intracellular parasites.
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Dependovirus /Virusoids 卫星病毒
Viroids are small (200-400nt), circular RNA molecules with a rod-like secondary structure which possess no capsid or envelope which are associated with certain plant diseases. Their replication strategy like that of viruses - they are obligate intracellular parasites.
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朊粒 (Prions)
Prions are rather ill-defined infectious agents believed to consist of a single type of protein molecule with no nucleic acid component. Confusion arises from the fact that the prion protein & the gene which encodes it are also found in normal 'uninfected' cells. These agents are associated with diseases such as Creutzfeldt-Jakob disease in humans, scrapie in sheep & bovine spongiform encephalopathy (BSE) in cattle.