Pignatelli J, Jimenez M, Luque J, Rejas M T, Lavazza A, Rodriguez D
Department of Molecular and Cellular Biology, Centro Nacional de Biotecnologia, CSIC, C/Darwin 3, 28049 Madrid, Spain.
Virus Res. 2009 Jul;143(1):33-43. doi: 10.1016/j.virusres.2009.02.019. Epub 2009 Mar 20.
Toroviruses are emergent viruses, belonging to the Nidovirales order, that remain mostly ignored, despite they are able to infect different species of domestic animals and humans, causing enteric diseases and diarrhea. Thus far, only five variants of porcine torovirus (PToV) have been identified. In this report we describe the identification and partial characterization of a new strain of porcine torovirus (PToV-BRES) that was detected by RT-PCR in a swine faecal specimen from a farm in Brescia (Italy). The complete genes coding for the nucleocapsid (N), hemagglutinin-esterase (HE) and membrane (M) proteins were amplified, and sequence analysis showed that PToV-BRES is a new PToV strain that, based on the HE gene sequence, is phylogenetically related to P4 strain, that was up to now the only member of a distinct PToV lineage. The nucleocapsid protein from PToV-BRES was expressed in insect cells as a his-tagged protein, purified by affinity chromatography and used to develop an ELISA method to detect antibodies against PToV. This assay was evaluated using a serum collection including 45 samples from three commercial farms from Spain. High antibody prevalence against PToV was observed in the three farms, both in adult animals and in piglets, which could suggest that PToV might be endemic in Spanish porcine population. The ELISA method developed in this work could be useful in future epidemiological surveys about toroviruses.
环曲病毒是新兴病毒,属于尼多病毒目,尽管它们能够感染不同种类的家畜和人类,引发肠道疾病和腹泻,但大多仍被忽视。迄今为止,仅鉴定出五种猪环曲病毒(PToV)变体。在本报告中,我们描述了一株新型猪环曲病毒(PToV-BRES)的鉴定和部分特征,该病毒是通过逆转录聚合酶链反应(RT-PCR)在来自意大利布雷西亚一个农场的猪粪便样本中检测到的。编码核衣壳(N)、血凝素酯酶(HE)和膜(M)蛋白的完整基因被扩增,序列分析表明PToV-BRES是一种新的PToV毒株,基于HE基因序列,它在系统发育上与P4毒株相关,而P4毒株是迄今为止一个独特的PToV谱系的唯一成员。来自PToV-BRES的核衣壳蛋白在昆虫细胞中作为带组氨酸标签的蛋白表达,通过亲和层析纯化,并用于开发一种检测抗PToV抗体的酶联免疫吸附测定(ELISA)方法。使用来自西班牙三个商业农场的45份血清样本对该检测方法进行了评估。在这三个农场的成年动物和仔猪中均观察到较高的抗PToV抗体流行率,这可能表明PToV在西班牙猪群中可能是地方性流行。本研究中开发的ELISA方法可能对未来关于环曲病毒的流行病学调查有用。