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High-throughput Detection Method for Influenza Virus
Published on: February 4, 2012
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Enterovirus subtyping in a routine UK laboratory setting between 2013 and 2017
Hannah C Howson-Wells1, Stephen Winckles2, Camille Aliker2
1Clinical Microbiology, Nottingham University Hospitals NHS Trust, Nottingham, NG7 2UH, United Kingdom.
Summary
A new RT-PCR assay offers improved human enterovirus (EV) genotyping, addressing significant typing failures in national reference laboratories. This enhanced method successfully identified previously untypable EV strains, improving diagnostic accuracy for viral meningitis.
Area of Science:
- Virology
- Molecular Biology
- Public Health
Background:
- Human enteroviruses (EV) are a primary cause of viral meningitis.
- Current EV genotyping relies heavily on VP1 gene sequencing by national reference laboratories (NRLs).
Purpose of the Study:
- To assess the frequency of EV genotyping failures in NRL-submitted samples.
- To implement and validate a superior alternative assay for untyped specimens.
Main Methods:
- Audited NRL genotyping data from 2013-2017 for UK patients.
- Identified and evaluated an alternative RT-PCR VP1-typing assay.
- Performed in silico analysis of primers against 2,201 EV genomes.
- Applied the optimized assay to previously untypable archived samples.
Main Results:
- A significant increase in untypable EV strains was observed over the study period.
- In silico analysis revealed primer mismatches in the reference assay for clinically significant EV species.
- The alternative RT-PCR assay successfully genotyped 23 of 24 previously untypable specimens (EV-A and EV-B).
Conclusions:
- A modified, highly sensitive RT-PCR assay is a suitable alternative to the current national reference VP1-typing assay.
- This improved assay is recommended for settings experiencing enterovirus typing failures.
