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) Oral, genital, neonatal, & ocular herpes, HSV
Varicella-zoster virus (VZV) encephalitis
Epstein-Barr virus (EBV) Chicken pox (varicella), shingles (zoster)
Cytomegalovirus (CMV) Infectious mononucleosis
Human herpesviruses 68 Infections in newborns & immunocompromised
Roseola, Kaposi sarcoma
Papillomavirus Human pappiloma virus (HPV) Warts, including genital warts that are linked to
cervical cancer
Parvovirus Parvovirus B-19 Fifth disease (erythema infectiosum)

Poxvirus Variola Smallpox

HUMAN RNA VIRUS

Common Family Name Representative Viruses Infection(s)
Arenaviruses Lymphocyte choriomeningitis virus (LCM) Aseptic meningitis or meningoencephalitis
Lassa virus Lassa fever
Astroviruses Astrovirus Gastroenteritis in children.
Bunyavirus Arbovirus (California encephalitis, La Crosse Encephalitis, hepatitis
virus)
Hantavirus (Sin Nombre virus) Hantavirus pulmonary syndrome (HPS)
Caliciviruses Norovirus Most common cause of infectious gastroenteritis in
US
Coronavirus Coronavirus Severe acute respiratory syndrome (SARS), cold-
like infections, pediatric diarrhea
Filovirus Marburg & Ebola viruses Hemorrhagic fever

Flavivirus Arboviruses (yellow fever virus, St. Louis Yellow fever, St. Louis encephalitis, dengue fever,
encephalitis virus, dengue virus, West West Nile virus infection
Nile virus)
Hepatitis C virus (HCV) Hepatitis C
Orthomyxoviruses Influenza A, B, & C Influenza
Paramyxovirus Measles virus Measles (rubeola)
Mumps virus Mumps
Parainfluenza virus RTI in children
Respiratory syncytial virus (RSV) RTI in infants, elderly, immunocompromised
Human metapneumovirus (HMPV) RTI
Picornavirus Enteroviruses (polioviruses, oxsackieviruses Polio, hand-foot-mouth disease, aseptic
A & B, echoviruses, enteroviruses) meningitis, others
Hepatitis A virus (HAV) Hepatitis A
Rhinovirus Common cold
Reovirus Rotavirus Most common cause of gastroenteritis in infants
& children
Retrovirus Human immunodeficiency viruses (HIV-1, AIDS
HIV-2)
Human T-lymphotropic viruses (HTLV-1, T-cell leukemia & lymphoma, tropical spastic
HTLV-2) paraparesis
Rhabdovirus Rabies virus Rabies
Togavirus Rubella virus Rubella (German measles)
Eastern, Western, & Venezuelan equine Eastern, Western, & Venezuelan encephalitis
encephalitis viruses
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 HSV, HPV
biopsy
GI tract Stool, rectal swab Adults: noroviruses, adenoviruses, enteroviruses.
Infants/children: rotavirus, adenoviruses

Respiratory tract Nasal aspirate, throat swab, nasopharyngeal Influenza A & B, parainfluenza virus, adenoviruses,
swab, bronchoalveolar lavage, lung biopsy RSV, HMPV, rhinovirus, enteroviruses

Skin Vesicle fluid or scrapings HSV, VZV, measles, rubella, enterovirus, parvovirus
B19
Urinary tract Urine Adenovirus, HSV, CMV

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

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

Swabs
alginate, cotton, wood are inhibitory for some viruses.

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

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

Deliver immediately. If not possible, keep at 28C & deliver within 2 hr.
Transport
Exception: Keep whole blood at room temp.

Best to process upon arrival. If not possible, hold at 28C for up to 48 hr. >48
Storage 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.
Shell vial culture 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
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.
Serology 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 EXAMPLE FOR ISOLATION OF
Cells from mammalian tissue Primary monkey kidney (PMK) Influenza viruses,
parainfluenza viruses,
enteroviruses
Primary
Rabbit kidney Herpes simplex virus
Human embryonic kidney Adenovirus, enteroviruses
Cultures of connective tissue Human neonatal lung (HNL) CMV (only type of cell
cells culture that can be used)
Finite (diploid)

Continuous (immortal,
heteroploid)
No single cell type grows all viruses. Several types should be inoculated.
Fibroblasts (e.g. MRC-5, WI-38) VZV, HSV, rhinovirus,
enteroviruses (some),
adenovirus, RSV
HEp-2 RSV, adenovirus, HSV,
parainfluenza viruses (some),
enteroviruses (some)
A549 HSV, adenovirus,
Malignant or transformed
enteroviruses
cells
MDCK Influenza viruses
LLC-MK2 Parainfluenza viruses, hMPV
Rhabdosarcoma (RD) Echovirus
Buffalo green monkey 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!