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使用快速抗原检测试剂盒、逆转录-聚合酶链反应(rt-PCR)和下一代DNA测序技术检测牛A组轮状病毒

Detection of bovine group a rotavirus using rapid antigen detection kits, rt-PCR and next-generation DNA sequencing.

作者信息

Minami-Fukuda Fujiko, Nagai Makoto, Takai Hikaru, Murakami Toshiaki, Ozawa Tadashi, Tsuchiaka Shinobu, Okazaki Sachiko, Katayama Yukie, Oba Mami, Nishiura Naomi, Sassa Yukiko, Omatsu Tsutomu, Furuya Tetsuya, Koyama Satoshi, Shirai Junsuke, Tsunemitsu Hiroshi, Fujii Yoshiki, Katayama Kazuhiko, Mizutani Tetsuya

机构信息

Ishikawa Nanbu Livestock Hygiene Service Center, Saida, Kanazawa, Ishikawa 920-3101, Japan.

出版信息

J Vet Med Sci. 2013 Dec 30;75(12):1651-5. doi: 10.1292/jvms.13-0265. Epub 2013 Aug 2.

Abstract

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.

摘要

我们研究了人轮状病毒快速抗原检测(RAD)试剂盒、逆转录聚合酶链反应(RT-PCR)和新一代DNA测序(NGS)检测牛A组轮状病毒(RVA)的敏感性。在有限稀释分析中,“荣研”试纸条法(Dipstick)在七种RAD试剂盒中对所有选定毒株的敏感性最高,这与先前报道的马轮状病毒检测结果一致。RT-PCR的敏感性比Dipstick高10⁰-10³倍。对13份RT-PCR阴性粪便样本进行NGS分析发现,所有样本均产生了RVA读数,其中两份样本覆盖了全部11个基因组片段。此外,将读数映射到参考序列可进行基因分型。NGS对于较少依赖特异性引物的分析和基因型筛选将是灵敏且有用的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ba3f/3942952/122cb34195d3/jvms-75-1651-g001.jpg

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