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Related Experiment Videos

Identification of enterically transmitted hepatitis virus particles by solid phase immune electron microscopy.

C D Humphrey1, E H Cook, D W Bradley

  • 1Ultrastructure Activity, Center for Infectious Diseases Centers for Disease Control, Atlanta, Georgia 30333.

Journal of Virological Methods
|August 1, 1990
PubMed
Summary

Identifying small enteric hepatitis viruses like hepatitis A virus (HAV) and hepatitis E virus (HEV) is challenging. Solid phase immune electron microscopy (SPIEM) significantly improves detection sensitivity and speed for these viruses in clinical samples.

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Area of Science:

  • Virology
  • Immunology
  • Microscopy

Background:

  • Small viruses (<50 nm), such as hepatitis A virus (HAV) and hepatitis E virus (HEV), are difficult to detect using standard immune electron microscopy (IEM).
  • Routine IEM methods struggle with identifying these viruses in complex biological matrices like stool and cell culture extracts due to interference from debris.

Purpose of the Study:

  • To evaluate the efficacy of a modified solid phase immune electron microscopy (SPIEM) technique for enhanced detection of small enteric hepatitis viruses.
  • To compare the sensitivity of SPIEM against conventional IEM for identifying hepatitis A virus (HAV) and hepatitis E virus (HEV).

Main Methods:

  • Utilized a combination of conventional immune electron microscopy (IEM) and a modified solid phase immune electron microscopy (SPIEM) methodology.

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  • Applied the techniques to identify HAV in stool and cell culture extracts, and HEV in stool extracts.
  • Investigated the effects of altered physical environments and potential modifications within the SPIEM assay.

    • Main Results:

    • The modified SPIEM method demonstrated a significant increase in the number of viral particles observed compared to conventional IEM.
    • SPIEM facilitated the visualization of small aggregates (2-20 particles) of HAV and HEV, which were not readily apparent with standard IEM.
    • The technique proved effective for both hepatitis A virus (HAV) and hepatitis E virus (HEV) detection in relevant clinical samples.

    Conclusions:

    • Solid phase immune electron microscopy (SPIEM) is a highly sensitive and specific assay for the rapid identification of enteric hepatitis viruses.
    • The SPIEM method offers a significant improvement over conventional IEM for detecting small, 'featureless' viruses in challenging sample types.
    • SPIEM facilitates faster and more accurate diagnosis of hepatitis A and E infections.