Rapid Serotyping of Enterovirus Infections

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Dr. Yu-Chieh Liao (廖玉潔)
Assistant Investigator
Institute of Population Health Sciences,
National Health Research Institutes,
Taiwan

ABSTRACT

Human enteroviruses are currently classified into 4 species including more than 100 serotypes. There are about 10 serotypes frequently related to severe complications. Therefore, it is necessary to develop high-throughput diagnostic tools for rapid serotyping of enterovirus infections using clinical specimens, which could be used for risk assessment to reduce unnecessary hospitalizations and class suspension. Recently, RT-PCR based on CODEHOP (Consensus-Degenerate Hybrid Oligonucleotide Primer) has been developed for targeting VP1 genes of enteroviruses (VP1-CODEHOP). The VP1-CODEHOP combining sequence analysis could successfully serotype enteroviruses using virus isolates and throat swabs. Moreover, the sequence-based VP1-CODEHOP has be applied to routine enterovirus surveillance in a medical center in Northern Taiwan (Chung WY et al. 2016 PLoS One). Therefore, the sequence-based VP1-CODEHOP could be further evaluated in National Enterovirus Surveillance System in Taiwan. Since the sequence-based VP1-CODEHOP requires 2-3 days and highly-trained technicians to complete the test, it is desirable to further develop user-friendly tools for rapid serotyping of enteovirus infections. Therefore, we are developing chip-based VP1-CODEHOP to use DNA probes for detecting serotype-specific nucleotide in VP1-CODEHOP derived PCR products. We first used bioinformatic modelling to identify potential serotype-specific probes. Then, the potential DNA probes were fixed in chips which were further reacted with VP1-CODEHOP derived PCR products labelled with biotin. The results could be read using naked eyes or machines and the overall time for completing test could be reduced to 24 hours. Currently, we has identified serotype-specific probes for 8 enterovirus serotypes. After probe optimization, DNA chips based on these serotype-specific probes could successfully identify these 8 serotypes. Detailed results will be presented in the presentation.

The content is authorized by Dr. Yu-Chieh Liao on October 30, 2017.