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首页> 外文期刊>Archives of virology >Detection and identification of avian, duck, and goose reoviruses by RT-PCR: goose and duck reoviruses are part of the same genogroup in the genus Orthoreovirus.
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Detection and identification of avian, duck, and goose reoviruses by RT-PCR: goose and duck reoviruses are part of the same genogroup in the genus Orthoreovirus.

机译：通过RT-PCR检测和鉴定禽，鸭和鹅呼肠孤病毒：鹅和鸭呼肠孤病毒是正咽病毒属同一基因组的一部分。

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A reverse transcription-polymerase chain reaction (RT-PCR) procedure for the detection of avian, duck, and goose reovirus (ARV, DRV, and GRV) RNA from cell culture supernatant and clinical samples was established. Based on multiple sequence alignment, a pair of degenerate primers was selected and synthesized. The amplified, cloned, and sequenced 598-base-pair products from the sigmaA-encoding gene fragment from 16 isolates (ranging over 30 years) indicated that the primer regions were well conserved. The sensitivity of this method was determined to be 10(-2) PFU. The specificity of the RT-PCR method was determined by testing specimens containing avian influenza A viruses, Newcastle disease virus, and infectious bronchitis virus, all of which yielded negative results with no discernible background. The efficiency of the system for detection of ARV, DRV, and GRV directly in 71/83 clinical samples was confirmed. The nucleotide sequence analysis indicated that DRV and GRV isolated from China in different locales and years were closely related, showing 97.4-100% homology to each other, but with only 86.7-88.5% identity to DRV 89026. The nucleotide and amino acid sequence identities in the amplified sigmaA-encoding gene were 74.2-78.4% and 86.9-92.0%, respectively, between duck/goose and chicken species. Phylogenetic analysis indicated that GRV and DRV aggregated into the same specified genogroup within subgroup II of the genus Orthoreovirus and are more closely related to ARV than to Nelson Bay virus. Overall, this study developed a sensitive and specific technique for the identification ARV, DRV, and GRV, and sequencing analysis has enhanced our understanding of the evolutionary relationship between ARV, DRV, and GRV.
机译：建立了从细胞培养上清液和临床样品中检测禽，鸭和鹅呼肠孤病毒（ARV，DRV和GRV）RNA的逆转录聚合酶链反应（RT-PCR）程序。基于多重序列比对，选择并合成了一对简并引物。来自16个分离株（范围超过30年）的sigmaA编码基因片段的扩增，克隆和测序的598个碱基对产物表明，引物区非常保守。该方法的灵敏度确定为10（-2）PFU。 RT-PCR方法的特异性是通过检测包含禽流感A病毒，新城疫病毒和传染性支气管炎病毒的标本来确定的，所有这些标本均产生阴性结果，而背景却不明显。证实了直接检测71/83临床样本中的ARV，DRV和GRV的系统的效率。核苷酸序列分析表明，在不同地区和年份从中国分离的DRV和GRV密切相关，彼此同源性为97.4-100％，但与DRV 89026的同源性仅为86.7-88.5％。核苷酸和氨基酸序列同一性鸭/鹅与鸡之间的编码sigmaA编码基因中的Aβ分别为74.2-78.4％和86.9-92.0％。系统发育分析表明，GRV和DRV聚集在正咽病毒属第二亚组内相同的特定基因组中，与ARV的关系比与尼尔森湾病毒的关系更密切。总体而言，这项研究开发了一种灵敏且特异的技术来鉴定ARV，DRV和GRV，而测序分析则使我们对ARV，DRV和GRV之间的进化关系有了更深入的了解。

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