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Identification of human picornaviruses by nucleic acid probes
1Department of Virology, University of Turku, Finland.
Abstract:
Human picornaviruses include rhinoviruses and enteroviruses which are responsible for both common and severe clinical diseases. Rhinoviruses are a frequent cause of respiratory infections while members of enterovirus subgroups, polio, coxsackie and ECHO viruses are often responsible for infections of the central nervous system, myocarditis, myositis etc. Human picornaviruses consist of nearly two hundred serotypes and therefore their specific identification after virus isolation, or the diagnosis based on the detection of immune response in patients, is problematic and does not usually provide virological diagnosis at the acute phase of illness. New methods for detection of picornavirus genomic RNA together with increasing knowledge of the nucleotide sequences of this virus group offer interesting possibilities for diagnostic procedures. Spot hybridization, in situ hybridization and enzymatic amplification of specific sequences have successfully been used for this purpose. Probes covering the 5' non-coding part of the genome, and also sequences derived from the region coding for non-structural proteins, can be used as broadly reacting reagents in picornavirus detection. Specific sequences are mainly found in the capsid protein region of the genome. cDNA probes and synthetic oligonucleotides are useful in rapid identification of picornaviruses after amplification in cell cultures and in epidemiological analysis. The biochemical amplification methods may enable recognition of picornaviruses directly in clinical samples in the near future. In situ hybridization methods have been of special interest because they can be used to reveal the presence of enterovirus genomes in biopsy specimens from e.g. affected heart muscle in patients with myocarditis and cardiomyopathy.
Insights
Detecting human picornaviruses like rhinoviruses and enteroviruses is challenging due to their many serotypes. New molecular methods offer improved diagnostic capabilities for these common and severe viral infections.
Area of Science:
- Virology
- Molecular Biology
- Infectious Diseases
Background:
- Human picornaviruses encompass rhinoviruses and enteroviruses, causing diverse illnesses from common colds to severe conditions like CNS infections and myocarditis.
- The large number of picornavirus serotypes complicates traditional identification methods, often delaying diagnosis during the acute phase of illness.
Purpose of the Study:
- To explore novel molecular diagnostic approaches for human picornaviruses.
- To evaluate the utility of genomic RNA detection and sequence knowledge for improved diagnostic procedures.
Main Methods:
- Utilized spot hybridization, in situ hybridization, and enzymatic amplification techniques for picornavirus detection.
- Employed probes targeting the 5' non-coding region and non-structural protein-coding sequences for broad reactivity.
- Investigated cDNA probes and synthetic oligonucleotides for rapid identification and epidemiological analysis.
Main Results:
- Demonstrated the successful application of hybridization and amplification methods for picornavirus detection.
- Identified conserved regions, including the capsid protein gene, suitable for broadly reactive probes.
- Showcased the potential of in situ hybridization for detecting enterovirus genomes in affected tissues, such as heart muscle biopsies.
Conclusions:
- Molecular methods, particularly those targeting viral genomic RNA, offer significant advantages over traditional techniques for picornavirus diagnosis.
- Enzymatic amplification holds promise for direct detection of picornaviruses in clinical samples in the near future.
- In situ hybridization is a valuable tool for diagnosing enterovirus-associated conditions like myocarditis by visualizing viral presence in tissue.