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Title: Evaluation of a reverse line blot assay for genotyping common human rotaviruses
Authors: Duizer, Erwin ×
van der Veer, Bas
de Bruin, Erwin
Zeller, Mark
Heylen, Elisabeth
Van Ranst, Marc
Matthijnssens, Jelle
Koopmans, Marion
Vennema, Harry #
Issue Date: Nov-2013
Publisher: Elsevier/North-Holland Biomedical Press
Series Title: Journal of Virological Methods vol:193 issue:2 pages:597-602
Article number: S0166-0934(13)00327-3
Abstract: To allow high-throughput genotyping of group A rotaviruses (RVA) in a routine surveillance setting, we developed a reverse line blotting method for the determination of the most common human RVA G- and P-genotypes: G1-G4, G9, G12, P[4], P[6] and P[8]. Using the reverse line blotting method on 951 clinical RVA positive feces samples, in 905 (95%) of the samples the G-genotyping yielded a result while in 945 (99%) of the samples the P-genotyping was successful. Comparison of the reverse line blotting-method as it is used currently to a sequence based method for genotyping RVAs showed an agreement of 96% for single strain infections (75 out of 78) but only 48% for mixed infections (10 out of 21). The reverse line blotting method is successful in genotyping common RVA strains in surveillance settings. For genotyping of rare strains, the number of probes on the blot can be expanded or a sequence-based method can be performed as a complementary approach on the samples that are positive in the detection PCR but negative in the reverse line blotting genotyping assay. The complete reverse line blotting protocol takes 8h to complete for 36 samples, with 3h hands-on-time. In conclusion, a RVA genotyping method that is accurate, cheap and fast is presented.
URI: 
ISSN: 0166-0934
Publication status: published
KU Leuven publication type: IT
Appears in Collections:Laboratory of Clinical and Epidemiological Virology (Rega Institute)
× corresponding author
# (joint) last author

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