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VIROLOGY REVIEW

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

Adenovirus serotypes 1-489

Respiratory, urinary tract, GI, & eye infections

Hepadnavirus

Hepatitis virus B (HBV)

Hepatitis

Herpes virus

Herpes simplex viruses (HSV-1, HSV-2)


Varicella-zoster virus (VZV)
Epstein-Barr virus (EBV)
Cytomegalovirus (CMV)
Human herpesviruses 68

Oral, genital, neonatal, & ocular herpes, HSV


encephalitis
Chicken pox (varicella), shingles (zoster)
Infectious mononucleosis
Infections in newborns & immunocompromised
Roseola, Kaposi sarcoma

Papillomavirus

Human pappiloma virus (HPV)

Parvovirus

Parvovirus B-19

Warts, including genital warts that are linked to


cervical cancer
Fifth disease (erythema infectiosum)

Poxvirus

Variola

Smallpox

HUMAN RNA VIRUS


Common Family Name

Representative Viruses

Infection(s)

Arenaviruses

Lymphocyte choriomeningitis virus (LCM)


Lassa virus

Aseptic meningitis or meningoencephalitis


Lassa fever

Astroviruses

Astrovirus

Gastroenteritis in children.

Bunyavirus

Arbovirus (California encephalitis, La Crosse


virus)
Hantavirus (Sin Nombre virus)

Encephalitis, hepatitis

Caliciviruses

Norovirus

Coronavirus

Coronavirus

Filovirus

Marburg & Ebola viruses

Most common cause of infectious gastroenteritis in


US
Severe acute respiratory syndrome (SARS), coldlike infections, pediatric diarrhea
Hemorrhagic fever

Flavivirus

Arboviruses (yellow fever virus, St. Louis


encephalitis virus, dengue virus, West
Nile virus)
Hepatitis C virus (HCV)
Influenza A, B, & C
Measles virus
Mumps virus
Parainfluenza virus
Respiratory syncytial virus (RSV)
Human metapneumovirus (HMPV)
Enteroviruses (polioviruses, oxsackieviruses
A & B, echoviruses, enteroviruses)
Hepatitis A virus (HAV)
Rhinovirus
Rotavirus

Orthomyxoviruses
Paramyxovirus

Picornavirus

Reovirus
Retrovirus

Rhabdovirus
Togavirus

Human immunodeficiency viruses (HIV-1,


HIV-2)
Human T-lymphotropic viruses (HTLV-1,
HTLV-2)
Rabies virus
Rubella virus
Eastern, Western, & Venezuelan equine
encephalitis viruses

Hantavirus pulmonary syndrome (HPS)

Yellow fever, St. Louis encephalitis, dengue fever,


West Nile virus infection
Hepatitis C
Influenza
Measles (rubeola)
Mumps
RTI in children
RTI in infants, elderly, immunocompromised
RTI
Polio, hand-foot-mouth disease, aseptic
meningitis, others
Hepatitis A
Common cold
Most common cause of gastroenteritis in infants
& children
AIDS
T-cell leukemia & lymphoma, tropical spastic
paraparesis
Rabies
Rubella (German measles)
Eastern, Western, & Venezuelan encephalitis

COMMON VIRUSES BY SITE


SITE OF INFECTION

SPECIMENS

COMMON VIRUSES

Centro nervous system

CSF, throat swab, stool, brain tissue, blood

Enterovirus, HSV, arbovirus

Eye

Conjunctival swab, corneal scraping

HSV, adenoviruses

Genital tract

Genital swab, vesicle swab or fluid, lesion


biopsy
Stool, rectal swab

HSV, HPV

Respiratory tract

Nasal aspirate, throat swab, nasopharyngeal


swab, bronchoalveolar lavage, lung biopsy

Influenza A & B, parainfluenza virus, adenoviruses,


RSV, HMPV, rhinovirus, enteroviruses

Skin

Vesicle fluid or scrapings

HSV, VZV, measles, rubella, enterovirus, parvovirus


B19

Urinary tract

Urine

Adenovirus, HSV, CMV

GI tract

Adults: noroviruses, adenoviruses, enteroviruses.


Infants/children: rotavirus, adenoviruses

VIRAL SPECIMEN TRANSPORT AND COLLECTION


Time of collection

During acute phase (1st 3-5 days)

Site of collection

Site of infection, entry & exit sites.

Collection containers

Sterile, leak-proof, non-breakable

Swabs

Dacron, rayon, or other polyester tips. Plastic or aluminum shafts. Calcium


alginate, cotton, wood are inhibitory for some viruses.

Transport media

Viral transport medium (VTM), 2-sucrose phosphate (2-SP), Amies or Stuarts


transport media. Not required for blood, CSF, urine.

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.)

METHODS FOR DIAGNOSIS OF VIRAL INFECTIONS


Cytology/histology

Microscopic examination of specimen for viral cytopathic effect (CPE).

Electron microscopy

Rarely used. Labor intensive, expensive.

Direct fluorescent antibody stain

Fluorescent-labeled antibody added to patient cells fixed to slide. If viral antigen


present, antibody binds. Fluorescence seen with fluorescent microscope.

Antigen detection

Solid-phase & membrane ELISAs.

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.

Shell vial culture

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)

Herpes simplex virus


Adenovirus, enteroviruses
CMV (only type of cell
culture that can be used)

Primary

Finite (diploid)

Cultures of connective tissue


cells

FOR ISOLATION OF

Fibroblasts (e.g. MRC-5, WI-38)

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.

VZV, HSV, rhinovirus,


enteroviruses (some),
adenovirus, RSV
RSV, adenovirus, HSV,
parainfluenza viruses (some),
enteroviruses (some)
HSV, adenovirus,
enteroviruses
Influenza viruses
Parainfluenza viruses, hMPV
Echovirus
Coxsackieviruses

Cytologic findings suggestive of viral infection:

Warthin-Finkeldey cells Measles

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

widely used to detect cell-associated viral antigens.

main applications is to detect respiratory, ocular, cutaneous, and bloodstream pathogens

major limitation is having an adequate number of cells in the specimen


EIA

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

NOMENCLATURE AND DEFINITIONS OF HEPATITIS VIRUSES ANTIGENS AND ANTIBODIES

AIDS AND LENTIVIRUS


HIV

derived from primate lentivirus


illness (AIDS) first described in 1981
isolated in 1983

Properties of HIV:

retrovirus, member of the family of Lentivirus genus

unique morphologic characteristic of HIV is a cylindrical nucleoid in the mature virion

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

Major gene products of HIV that are useful in Dx of Infection

antibodies to viral proteins of gp120, gp41, p24 are commonly detected

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

Test kits commercially available for measuring antibodies by EIA

Most widely used confirmatory assay Western Blot

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

Ad majorem, Dei gloriam!