Beikpour Farzad, Pellegrini Francesco, Lanave Gianvito, Camero Michele, Catella Cristiana, Di Martino Barbara, Di Profio Federica, Masotti Chiara, Battistini Roberta, Serracca Laura, La Rosa Giuseppina, Martella Vito, Suffredini Elisabetta
Department of Veterinary Medicine, University of Bari Aldo Moro, 70010 Valenzano, Italy.
Faculty of Veterinary Sciences, University of Teramo, Località Piano d'Accio, 64100 Teramo, Italy.
Vet Sci. 2023 Feb 21;10(3):175. doi: 10.3390/vetsci10030175.
Astroviruses are important human enteric pathogens transmissible with contaminated food and water. Astroviruses have also been identified in mammals, birds, lower vertebrates and invertebrates. The genetic diversity of human and animal astroviruses poses a challenge for diagnostics and taxonomy. As a proof of concept, we used a panastrovirus consensus primer set, able to amplify in a nested RT-PCR protocol a 400-nt-long fragment of the RNA-dependent RNA polymerase of most members of the family, in conjunction with a nanopore sequencing platform, to generate information on the astrovirome in filter-feeding mollusks. Amplicons generated from bivalve samples were used to generate libraries for deep sequencing. In three samples, only one unique RdRp sequence type was obtained. However, in seven samples and in three barcodes with eleven pooled samples, we identified a variety of known and unknown RdRp sequence types, in most cases distantly related to astrovirus sequences available in the databases. In total, 37 different sequence contigs were generated. Avian-origin astrovirus sequences were predominant, likely due to contamination of shellfish harvesting waters by marine birds. Astroviruses of the aquatic eco-system were also identified, whereas human astroviruses were not detected.
星状病毒是重要的人类肠道病原体，可通过受污染的食物和水传播。星状病毒也已在哺乳动物、鸟类、低等脊椎动物和无脊椎动物中被发现。人类和动物星状病毒的遗传多样性对诊断和分类学构成了挑战。作为概念验证，我们使用了一套泛星状病毒通用引物，该引物能够在巢式逆转录聚合酶链反应（RT-PCR）方案中扩增该病毒家族大多数成员的依赖RNA的RNA聚合酶的一段400个核苷酸长的片段，并结合纳米孔测序平台，以获取滤食性软体动物中星状病毒组的信息。从双壳贝类样本中产生的扩增子用于生成深度测序文库。在三个样本中，仅获得了一种独特的RNA依赖的RNA聚合酶（RdRp）序列类型。然而，在七个样本以及三个包含11个混合样本的条形码中，我们鉴定出了多种已知和未知的RdRp序列类型，在大多数情况下，这些序列与数据库中可用的星状病毒序列关系较远。总共产生了37种不同的序列重叠群。禽源星状病毒序列占主导地位，这可能是由于海鸟对贝类捕捞水域的污染所致。水生生态系统中的星状病毒也被鉴定出来，而未检测到人类星状病毒。
