Sie sind auf Seite 1von 12

Lab Diagnosis of Viral Diseases

S. Saengamnatdej, PhD.
for 499305 (2009/2)
April 25, 2009
Goals
Specimen Processing
Collection of Specimens
Methods
Goals
1. To confirm the diagnosis (acute or chronic infection).
2. To determine appropriate antiviral therapy.
3. To define the course of disease.
4. To determine the immune status.
5. To establish public health measures to prevent the spread.
6. To study the evolution & epidemiology of viral infections.
Specimen Flow Diagram
Request Report
Macroscopic & Microscopic
Transport Examination Initial (via phone)

Collect Antigen detection;


nucleic acid probes
Preliminary
(same day)

Preliminary
Cultivation, isolation Identification
(day after receipt)
viruses, fungi &some bacteria
may take longer time.

Identification;
antimicrobial sensitivity testing Final (2nd day)

Further identification;
Supplementary
typing; toxin production;
Slack, R.C.B. (2006) further sensitivities
Collection of Specimens

- Avoid contamination
(commensals or external sources)

- Use appropriate methods.

- In a transport-durable, leak-proof,
sterile container.

- Use viral transport medium


containing albumin and suitable antibiotics.
Specimens

- Sent with a request card.


(providing sufficient information.)

- Labeled correctly.
- Kept separately from food and drug.
- Transported on ice (4 ºC) without delay.
(frozen, if undelivered for weeks.)

- Follow safety regulation.


(each step with a responsible personnel and minimize handling by
untrained people.)
Methods of Collection
Eye Conjunctival swab

Respiratory secretions Nasopharyngeal swab, Throat


Adenovirus, Influenza, Enterovirus, (pharyngeal) swab, Nasal washing,
Rhinovirus, Paramyxovirus, Rubella, HSV Nasal swab
(ex: meningitis during summer—enterovirus)

GI specimens Vomitus,
Reovirus, Rotavirus, Stool, Rectal swab (meningitis during
Adenovirus, Norwalk, Calicivirus summer— enterovirus)
Methods of Specimen Collection (Cont.)

CNS CSF
(ex: CNS disease after parotitis—mumps
or meningitis during summer—enterovirus)
Brain biopsy: rabies

Genital Ulcer scrape

Blood Serum antigen, Serum antibody


HIV, HTLV, HBV,
HCV, HDV

Urine
Rubella, Measles, Adenovirus, CMV,
Mumps (ex: CNS disease after parotitis, Maculopapular rash)
Timing : Viral shedding
For examples,

Respiratory viruses
shed for only 3-7 days

HSV & VZV from lesions:


more than 5 days after the onset.

Enteroviruses from CSF ,


only 2-3 days after the onset.
1.Detect viral components Methods
Proteins
Nucleic acids

2.Serology for hard-to-isolate&grow, slow-disease causing v.


EBV, Rubella, Hepatitis A-E, HIV, HTLV, Arbovirus

3.Cytology
CPEs

4.Cultivation & Isolation


Animals/ Embryonated eggs/ Organ culture
/ Tissue culture (primary, diploid, continuous cell lines)
CPEs, plaques, interference, hemadsorption

5.Electron Microscopy
Notes on Serology Methods.

To indicate seroconversion, the antibody titre should


increase at least fourfold.
Why fourfold?
Because twofold increase in the result may be not
correct due to twofold serial dilutions in
the serologic assays have the imprecision nature.

Let's take these figures as example,


“samples with 512 and 1023 units of antibody would both
give a signal on a 512-fold dilution but not on a 1024-fold
dilution, and the titers of both would be reported as 512. On
the other hand, samples with 1020 and 1030 units are not
significantly different but would be reported as titers of 512
and 1024, respectively.”
(Murray, P.K. et al.,2005)
Notes on Serology Methods.(2)

How does serology help to define


the stage of diseases?

The antibody against the intracellular viral component


indicates the late stage of the infection.

For example, the antibody to EBV-nuclear antigen is


detected late in the infection.

(Murray, P.K. et al.,2005)