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Updated: Jul 27, 2026
Particle Agglutination Method for Poliovirus Identification
Published on: April 20, 2011
A novel serological technique: polymerase chain reaction enhanced immunoassay. Application to enterovirus IgM
Insights
A novel PCR enhanced immunoassay (PIA) detects antiviral antibodies by combining serology with PCR. This sensitive method shows promise for diagnosing viral infections, including enterovirus IgM.
Area of Science:
- Virology
- Immunology
- Molecular Biology
Background:
- The polymerase chain reaction (PCR) is a sensitive technique for virus detection.
- A PCR enhanced immunoassay (PIA) combines serological techniques with PCR for versatile antibody detection.
Purpose of the Study:
- To describe the principles and application of the PIA technique for antibody detection.
- To evaluate the PIA for detecting anti-enterovirus IgM in patient serum specimens.
Main Methods:
- The PIA method involves capturing IgM to antibody-coated plates, binding viral antigen, and using released RNA for reverse transcription PCR (RT-PCR).
- Amplicons are detected via hybridization to a labeled probe in a colorimetric assay.
- A pilot study examined 18 serum specimens from patients with enterovirus infections using a mixture of ten crude enterovirus antigens.
Main Results:
- The PIA detected IgM positivity in 6/18 (33%) of specimens, with titres ranging from 1/500 to 1/100,000.
- Predominantly type-specific antibodies were detected.
- Results were compared to a procapsid enterovirus radioimmunoassay (RIA).
Conclusions:
- The PIA offers new possibilities for virus serology, linking antigenicity with viral sequence.
- The PIA has potential as a clinically useful assay for detecting antiviral antibodies after further optimization.
Abstract:
The polymerase chain reaction (PCR) method is a sensitive, specific and rapid technique for virus detection. The principles of a PCR enhanced immunoassay (PIA) are described. The method combines solid phase serological techniques with the PCR, providing a versatile and sensitive method for antibody detection. By linking the antigenicity of virus particles with their content of nucleic acid, the method provides new possibilities for virus serology: for example, antibody specificity can be coupled to viral sequence in patients with chronic infections caused by highly variable viruses such as HIV and HCV. An application of the PIA technique is described for the detection of anti-enterovirus IgM. IgM is captured to anti-human IgM-coated microwell plates. The anti-enterovirus IgM is allowed to bind crude enterovirus antigen. Bound virus is heat denatured and the released RNA is used as a template for reverse transcription PCR (RT-PCR) amplification. Amplicons are detected by hybridisation to an affinity labelled probe in a microwell colorimetric assay. In a pilot study, 18 serum specimens from patients with enterovirus infections were examined. Using a mixture of ten crude enterovirus antigens, the frequency of IgM positivity was 6/18 (33%). Titres between 1/500 and 1/100,000 were recorded. Predominantly type-specific antibodies were detected. The results were compared with a procapsid enterovirus radioimmunoassay (RIA). After further optimisation, the PIA has the potential to be a clinically useful assay for the detection of antiviral antibodies.
