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Virion:
complete virus particle
Nucleocapsid:
nucleic acid & capsid
Nucleic acid:
DNA or RNA. Single- or double-stranded. Linear or circular.
Capsid:
Protein coat that encloses genetic material. May be helical (rod-like) or icosahedral (cuboid). Composed of protein
subunits called capsomers. Protects nucleic acid, enables virus to attach to & enter host cell.
Envelope: Outer membrane surrounding capsid in some viruses. Aids in attachment to host cell. Viruses without called naked
nucleocapsids.
Viral replication
Adsorption:
attachment of virus to host cell receptor.
Penetration:
Virus enters host cell by direct penetration, endocytosis (entering in a vacuole), or fusion with cell membrane.
Uncoating:
Loss of capsid. Genome enters cytoplasm most RNA viruses) or nucleus (most DNA viruses).
Eclipse/synthesis: Eclipse: several hours during which virions cant be detected. Synthesis: mRNA is produced. Directs synthesis of viral
particles.
Maturation/release:
Genetic material assembled into protein coat. Virions migrate to cytoplasmic membrane. Released by
budding off, leaking out, or lysing host cell with enzymes.
HUMAN DNA VIRUS
Common Family Name
Representative Viruses
Infection(s)
Adenovirus
Hepadnavirus
Hepatitis
Herpes virus
Papillomavirus
Parvovirus
Parvovirus B-19
Poxvirus
Variola
Smallpox
Representative Viruses
Infection(s)
Arenaviruses
Astroviruses
Astrovirus
Gastroenteritis in children.
Bunyavirus
Encephalitis, hepatitis
Caliciviruses
Norovirus
Coronavirus
Coronavirus
Filovirus
Flavivirus
Orthomyxoviruses
Paramyxovirus
Picornavirus
Reovirus
Retrovirus
Rhabdovirus
Togavirus
SPECIMENS
COMMON VIRUSES
Eye
HSV, adenoviruses
Genital tract
HSV, HPV
Respiratory tract
Skin
Urinary tract
Urine
GI tract
Site of collection
Collection containers
Swabs
Transport media
Transport
Deliver immediately. If not possible, keep at 28C & deliver within 2 hr.
Exception: Keep whole blood at room temp.
Storage
Best to process upon arrival. If not possible, hold at 28C for up to 48 hr. >48
hr, freeze at 70C.
(Not recommended.)
Electron microscopy
Antigen detection
Cell culture
Different viruses grow in different cell lines. Growth may take 128 days. Examine
microscopically for cytopathic effects (CPE): cell rounding, clumping, vacuolation,
granulation, giant multinucleate cells, cell fusion, syncytial formation, cell lysis,
plaques (groups of killed cells), inclusion bodies. Not all viruses produce CPE.
immunofluorescent stains may be used for confirmation.
Rapid modification of conventional cell culture. Detection in 12 days. Specimen
centrifuged onto monolayer of cells growing on coverslip. Coverslips stained with
viral-specific immunofluorescent conjugate. Used primarily for viruses that are slow to
produce CPE.
Molecular methods
Serology
PCR, real-time PCR, branched DNA, nucleic acid hybridization. Faster & more
sensitive than cell culture. Can detect viruses that cant be cultured, multiple viruses
simultaneously.
Detects antibodies in serum. Useful in evaluating immune status or diagnosing viral
infections where culture is difficult or impossible. Presence of antibodies isnt always
indicative of current infection.
CELL CULTURES
CELL LINE
DEERIVATION
Cells from mammalian tissue
EXAMPLE
Primary monkey kidney (PMK)
Influenza viruses,
parainfluenza viruses,
enteroviruses
Rabbit kidney
Human embryonic kidney
Human neonatal lung (HNL)
Primary
Finite (diploid)
FOR ISOLATION OF
HEp-2
Continuous (immortal,
heteroploid)
Malignant or transformed
cells
A549
MDCK
LLC-MK2
Rhabdosarcoma (RD)
Buffalo green monkey
No single cell type grows all viruses. Several types should be inoculated.
Cervical smear showing multinucleated cells and the Cowdry type A intranuclear inclusions herpes simplex infection
In paps smear, binucleate squamous epithelial cells with distinct perinuclear halos (koilocytosis) human papilloma virus
Urinary epithelial cell containing an enlarged nucleus with smudgy chromatin and a small pale glassy intranuclear inclusion
polyomavirus infection
Cell from bronchoalveolar lavage with a large intranuclear inclusion with a perinuclear clear space (owls eye cell) CMV
infection
Antigen detection
Fluorescent antibody staining
versatile and widely used method that can be applied to the detection of antigens, regardless of whether they are cell
associated or in fluid phase
Advantages include applicability to diverse specimens and potential for automation
HEPATITIS VIRUS
Hepatitis A: infectious hepatitis
Hepatitis B: serum hepatitis
Hepatitis C: common cause of post-transfusion hepatitis
Hepatitis D: defective virus dependent on coinfection
Hepatitis E: agent of enterically transmitted hepatitis
Characteristics of Hepatitis
Properties of HIV:
contain the four genes required for a replicating retrovirusgag, pro, pol, and envand follow the general pattern for
retrovirus replication
Early phase replication:
Tat protein functions in transactivation
Rev protein required for the expression of viral structural proteins
Nef protein increases viral infectivity, facilitates activation of resting T cells, and down regulates expression of CD4 and MHC class I
Vpr protein increases transport of the viral preintegration complex into the nucleus and also arrests cells in the G2 phase of the cell
cycle
Vpu protein promotes CD4 degradation
LABORATORY DIAGNOSIS
Detection in three ways:
1. virus isolation
2. serologic determination of antiviral antibodies
3. measurement of viral nucleic acid or antigens
PCR amplification techniques are commonly used for detection of virus in clinical specimens.
Serology
High titers of HIV are found in two body fluidsblood and semen
Ab against the Gag proteins (p17, p24, p55) appear earliest in the course of HIV-1infection, but decrease in titer with
progression of HIV-1 disease,
Antibodies to envelope (gp160 or gp120/gp41) usually persist even in advanced stages of HIV-1 disease
Prep. By: Terence Eday, RMT, MT(ASCPi), MPH for Minds Review Center