Diagnostic Methods in Virology

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Criteria for diagnosing Primary Infection
  1. A significant rise in titre of IgG/total antibody between acute and convalescent sera - however, a significant rise is very difficult to define and depends greatly on the assay used. In the case of CFT and HAI, it is normally taken as a four-fold or greater increase in titre. The main problem is that diagnosis is usually retrospective because by the time the convalescent serum is taken, the patient had probably recovered.

  2. Presence of IgM - EIA, RIA, and IF may be are used for the detection of IgM. This offers a rapid means of diagnosis. However, there are many problems with IgM assays, such as interference by rheumatoid factor, re-infection by the virus, and unexplained persistence of IgM years after the primary infection.

  3. Seroconversion - this is defined as changing from a previously antibody negative state to a positive state e.g. seroconversion against HIV following a needle-stick injury, or against rubella following contact with a known case.

  4. A single high titre of IgG (or total antibody) - this is a very unreliable means of serological diagnosis since the cut-off is very difficult to define.

Criteria for diagnosing re-infection/re-activation

It is often very difficult to differentiate re-infection/re-activation from a primary infection. Under most circumstances, it is not important to differentiate between a primary infection and re-infection. However, it is very important under certain situations, such as rubella infection in the first trimester of pregnancy: primary infection is associated with a high risk of fetal damage whereas re-infection is not. In general, a sharp large rise in antibody titres is found in re-infection whereas IgM is usually low or absent in cases of re-infection/re-activation.

Serological events following primary infection and reinfection. Note that in reinfection, IgM may be absent or only present transiently at a low level.

 
Overview
Direct Examination
- Electron Microscopy (EM)
- Light Microscopy
Virus Isolation
- Identification of growing virus
- Problems with cell culture
Serology
- Criteria for diagnosing
- Limitations of serological diagnosis
- Antibody in the CSF