Application of Molecular Biology in
Medical Virology – Specimen Collection,
Storage and Transport
Specimen Processing for Diagnosing
Viral Diseases
 Viruses are in highest
concentrations during the first
several days following onset of
symptoms. Therefore, samples
should be collected early in the
disease course.
 Viruses are fastidious: sensitive to
heat and drying
 Clinical specimens should be
handled with care and delivered to
the laboratory as soon as possible
 NEVER use Calcium Alginate
swabs: use DACRON or RAYON
SAMPLES:
1. Heparinized Whole Blood – for
viral / bacterial culture
2. EDTA Plasma – for Serology and
PCR
Samples should be generally come from the
infected site.
a. Skin Infections – rash site and
depending on the virus, serum or
urine
b. Respiratory infections – sputum or
throat swabs
c. Central Nervous System
(meningitis and encephalitis) – for
diagnosis of meningitis, CSF and
serum, as well as stool or throat
swabs, can be collected because
viruses are sometimes shed into
these sites. In case of encephalitis,
brain biopsy and sometimes serum
are used.
d. Urogenital infections – needle
aspirates and endocervical and
urethral swabs
e. Gastrointestinal tract – stool
samples and rectal swabs
f. Eye infections – eye swabs and
corneal scrapings
Sample Transport
Samples for viral culture must be placed
into a Viral Transport Medium (VTM).
VTM Contains:
a. Buffered Saline
b. Protein Stabilizers
c. Antimicrobials that inhibit bacterial
and fungal growth
 Samples for viral cultures can be
refrigerated in VTM for about 48
hours, they should never be frozen
at -20°C. Samples can be stored at -
70°C; however, infectivity will be
diminished.
Viral Isolation
1. CELL CULTURE
 Important means for diagnosing viral
infections
 Requires nutritionally rich complex
media
 The media often contains fecal calf
serum as a nutrient
 Some viruses have not yet been
grown in vitro.
 very sensitive. For these reasons,
electron microscopy is not
2. PRIMARY CELL CULTURES
commonly used.
 Derived directly from tissues
 Can only be maintained for a short
time inside the laboratory
Other methods of detection
 Example: Primary Monkey Kidney
Cells 1. Detection of host antibodies directed
against specific viruses
 Transferring cells from one container
to another is called “splitting” or 2. Direct detection of viral antigens in
“passaging” clinical specimens
3. ESTABLISHED CELL LINES 3. Viral gene probes and nucleic acid
amplification (e.g., polymerase chain
 Also referred to as low passage or
reaction [PCR])
finite cell lines
 Can be maintained longer than
primary cell lines
 Examples: WI – 38, MRC – 5, IMR -
90
4. CONTINUOUS CELL LINES
 Are altered cells that can be
maintained indefinitely
 Examples: HeLa Cells, HEp – 2,
A549, and Vero Cells
 They can be used to grow HSV,
VZV, CMV, Adenovirus and
Rhinoviruses.

Electron Microcopy
 Due to size, most individual virions
can only be seen by electron
microscopy.
 Electron Microscopy is sometimes
used to identify Norwalk viruses,
Astrovirus, Calicivirus, and
Coronavirus.
 Electron Microscope is expensive,
requires expertise and is usually