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Laboratory Diagnosis of Smallpox Virus
Laboratory Diagnosis of Smallpox Virus
B. LABORATORY DIAGNOSIS:
1. Sample Collection
2. Light Microscopy and Histopathology
exam of stained smears
3. Electron Microscopy of pustular fluid
4. Tissue Culture
5. Serological/Immunological Tests
a. ELISA
b. Immunofluroscence Test
c. Virus Neutralization Test
d. Complement Fixation Test
e. Heamagglutination-inhibition Test
6. Molecular Diagnostic Techniques
- PCR
A. Clinical Diagnosis
Initial diagnosis of a smallpox
infection is most likely based on a
history and physical examination
findings.
Smallpox is an illness
characterized by:
Severe headache
Backache
Fatigue
Malaise
Vomiting
Diarrhea
Excessive bleeding (in some cases)
** Before a diagnosis of smallpox is made, the doctor will consider other
illnesses that can mimic the signs and symptoms of smallpox.
Some of these illnesses include:
Chickenpox, Monkeypox, Herpes Zoster (Shingles), and Impetigo.
:Smallpox vs Chickenpox
a) Definition - Neutralization of a virus is defined as the “loss of infectivity through reaction of the virus
with its specific antibody”.
b) Purpose - Neutralization test is used to determine the antiviral activity of the serum of a suspected
case. (I.e.In this test, the serum of the patient is tested for its power to "neutralize" variola virus in tissue culture
or on CAM).
C) Test Steps - 1) Mixing the virus with the serum to be tested (which is supposed to contain virus-
neutralizing antibodies) under appropriate condition, and then 2) Inoculating that mixture into cell culture,
eggs or animals. Neutralization test, therefore, requires that an active suspension of that virus should be at
hand.
:Disadvantages of Neutralization Test**
b) The results are not always easy to interpret if the patient has been previously vaccinated against
smallpox. Neutralizing antibodies may persist in serum for years after smallpox vaccination, and usually
fade in about 10 years.
Antibodies against the viral protein responsible for haemagglutination can prevent
haemagglutination; this is the basis behind the haemagglutination-inhibition test
(HAI).
.(:Serological Tests (cont