Key result
RT-PCR demonstrates ~1000-fold higher sensitivity than rapid antigen kits, while NGS detects RT-PCR-negative rotavirus.
Next-generation sequencing is highly sensitive for detecting and genotyping bovine group A rotavirus, even in RT-PCR-negative samples.
NGS may enhance bovine rotavirus detection beyond RT-PCR in research; leaves open routine veterinary diagnostic adoption.
We investigated the sensitivity of human rotavirus rapid antigen detection (RAD) kits, RT-PCR and next-generation DNA sequencing (NGS) for detection of bovine group A rotavirus (RVA). The Dipstick 'Eiken' Rota (Dipstick) showed the highest sensitivity out of the seven RAD kits against all selected strains in limited dilution analyses, which was consistent with the results for equine rotavirus previously reported. RT-PCR had 10⁰-10³-fold higher sensitivity than the Dipstick. NGS using thirteen RT-PCR-negative fecal samples revealed that all samples yielded RVA reads and especially that two of them covered all 11 genome segments. Moreover, mapping reads to reference sequences allowed genotyping. The NGS would be sensitive and useful for analysis of less dependent on specific primers and screening of genotypes.
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Minami-Fukuda et al. (2013) studied Bovine group A rotavirus infection (n=13). Rapid antigen detection kits, RT-PCR, and next-generation DNA sequencing vs. Comparison among diagnostic methods was evaluated on Detection limit and sensitivity for bovine group A rotavirus. RT-PCR demonstrated up to 1000-fold higher sensitivity than the most sensitive rapid antigen detection kit, while next-generation sequencing successfully detected bovine rotavirus A in all 13 RT-PCR-negative fecal samples.
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