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Diagnosis of human parvovirus infection by dot-blot hybridization using cloned viral DNA
Abstract:
The human parvovirus can be detected in serum by the immunological techniques of immune electron microscopy (IEM), counterimmunoelectrophoresis (CIE), and radioimmunoassay (RIA). A portion of the genome of this virus has been cloned in pAT153 and used as a 32P-labelled probe in dot-blot hybridization assays to detect parvovirus DNA in serum specimens. This test proved a highly sensitive means of detecting virus in microlitre volumes of serum, giving positive results for samples containing 0.5 pg viral DNA, equivalent to 10(4) virus particles. Unlike CIE and RIA the test is not affected by the presence of parvovirus-specific antibody in serum specimens, and has permitted virus to be detected in specimens obtained up to 11 days after the onset of clinical symptoms of aplastic crisis.
Insights
A new dot-blot hybridization assay detects human parvovirus DNA in serum, offering high sensitivity. This method is unaffected by antibodies and aids in diagnosing aplastic crisis.
Area of Science:
- Virology
- Molecular Biology
- Immunology
Background:
- Human parvovirus is a significant human pathogen.
- Current detection methods include immune electron microscopy (IEM), counterimmunoelectrophoresis (CIE), and radioimmunoassay (RIA).
- These immunological techniques can be limited by the presence of specific antibodies in serum.
Purpose of the Study:
- To develop a highly sensitive method for detecting human parvovirus DNA in serum.
- To overcome limitations of existing immunological detection methods.
Main Methods:
- Cloning a portion of the human parvovirus genome into pAT153.
- Using the cloned DNA as a 32P-labelled probe for dot-blot hybridization assays.
- Detecting parvovirus DNA in microlitre volumes of serum specimens.
Main Results:
- The dot-blot hybridization assay demonstrated high sensitivity, detecting as little as 0.5 pg of viral DNA (equivalent to 10(4) virus particles).
- The test is not affected by the presence of parvovirus-specific antibodies in serum specimens.
- The assay successfully detected parvovirus DNA in samples collected up to 11 days after the onset of aplastic crisis symptoms.
Conclusions:
- Dot-blot hybridization using a cloned viral DNA probe is a sensitive and reliable method for detecting human parvovirus in serum.
- This technique offers an advantage over immunological methods by not being affected by antibodies.
- The assay facilitates early and accurate diagnosis of human parvovirus infections, particularly during aplastic crisis.