Detection of bovine group a rotavirus using rapid antigen detection kits, rt-PCR and next-generation DNA sequencing.
Detection of bovine group a rotavirus using rapid antigen detection kits, rt-PCR and next-generation DNA sequencing.
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DOI：
10.1292/jvms.13-0265
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发表时间：
2013-12-30
期刊：
影响因子：
--
通讯作者：
Mizutani T
中科院分区：
文献类型：
--
作者：
Minami-Fukuda F;Nagai M;Takai H;Murakami T;Ozawa T;Tsuchiaka S;Okazaki S;Katayama Y;Oba M;Nishiura N;Sassa Y;Omatsu T;Furuya T;Koyama S;Shirai J;Tsunemitsu H;Fujii Y;Katayama K;Mizutani T
研究了人轮状病毒快速抗原检测(RAD)试剂盒、RT-PCR和新一代DNA测序(NGS)试剂盒检测牛A组轮状病毒(RVA)的敏感性。在有限稀释分析中，在七种RAD试剂盒中，Diptick‘Eiken’ROTA(Diptick)对所有选定的毒株显示出最高的敏感性，这与先前报道的马轮状病毒的结果一致。RT-PCR的灵敏度是试纸条的100-103倍。使用13个RT-PCR阴性的粪便样本的NGS显示，所有样本都产生了RVA读数，特别是其中两个样本覆盖了全部11个基因组片段。此外，将读数映射到参考序列可以进行基因分型。NGS技术具有较高的灵敏度，可用于对特异性引物依赖性较低的分析和基因分型的筛选。
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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