Molecular detection and characterization of human enteroviruses directly from clinical samples using RT-PCR and DNA sequencing

Miren Iturriza-Gómara*, Brian Megson, Jim Gray

*Corresponding author for this work

    Research output: Contribution to journalArticlepeer-review

    94 Citations (Scopus)


    Enteroviruses are common human pathogens associated with a wide spectrum of symptoms ranging from asymptomatic infection to acute flaccid paralysis and neonatal multi-organ failure. Molecular methods that provide rapid diagnosis and increased sensitivity have been developed for the diagnosis of enterovirus infection using oligonucleotide primers complementary to conserved sequences located in the 5′ untranslated region (UTR), but data generated from these regions are not sufficiently discriminatory for typing due to the lack of correlation between their nucleic acid sequence and serotype specificity. Sequences derived from the gene encoding the capsid VP1 correlate with serotype, and therefore provide the opportunity for the development of molecular typing methods consistent with present serogical methods. In this study, oligonucleotide primers that amplify a region of the 5′UTR to detect enterovirus RNA, and the region encoding the enterovirus VP1 N-terminus to characterize virus strains were used in nested and semi-nested RT-PCRs, respectively. The ability of the VP1 RT-PCR to amplify diverse viruses within genotypes and genogroups was confirmed by the correct identification of both prototype strains, and strains circulating currently of the same genotypes. The mole-cular methods proved their utility through the detection of enteroviruses that failed to grow in cell culture, their subsequent characterization and the characterization of strains that failed to serotype in neutralization assays. Molecular methods increased significantly the sensitivity of detection (P < 0.001) and of characterization (P < 0.01) of enteroviruses when compared to classical methods.

    Original languageEnglish
    Pages (from-to)243-253
    Number of pages11
    JournalJournal of Medical Virology
    Issue number2
    Publication statusPublished - Feb 2006

    Bibliographical note

    Copyright 2008 Elsevier B.V., All rights reserved.


    • Enterovirus
    • Genotype
    • RT-PCR
    • VP1


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