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智利传染性胰腺坏死病毒的检测与系统发育分析

Detection and phylogenetic analysis of infectious pancreatic necrosis virus in Chile.

作者信息

Tapia D, Eissler Y, Torres P, Jorquera E, Espinoza J C, Kuznar J

机构信息

Centro de Investigación y Gestión de Recursos Naturales, Instituto de Química y Bioquímica, Facultad de Ciencias, Universidad de Valparaíso, Gran Bretaña 1111, Valparaíso 2360102, Chile.

出版信息

Dis Aquat Organ. 2015 Oct 27;116(3):173-84. doi: 10.3354/dao02912.

Abstract

Infectious pancreatic necrosis virus (IPNV) is the etiological agent of a highly contagious disease that is endemic to salmon farming in Chile and causes great economic losses to the industry. Here we compared different diagnostic methods to detect IPNV in field samples, including 3 real-time reverse transcription PCR (qRT-PCR) assays, cell culture isolation, and indirect fluorescent antibody test (IFAT). Additionally, we performed a phylogenetic analysis to investigate the genogroups prevailing in Chile, as well as their geographic distribution and virulence. The 3 qRT-PCR assays used primers that targeted regions of the VP2 and VP1 genes of the virus and were tested in 46 samples, presenting a fair agreement within their results. All samples were positive for at least 2 of the qRT-PCR assays, 29 were positive for cell culture, and 23 for IFAT, showing less sensitivity for these latter 2 methods. For the phylogenetic analysis, portions of 1180 and 523 bp of the VP2 region of segment A were amplified by RT-PCR, sequenced and compared with sequences from reference strains and from isolates reported by previous studies carried out in Chile. Most of the sequenced isolates belonged to genogroup 5 (European origin), and 5 were classified within genogroup 1 (American origin). Chilean isolates formed clusters within each of the genogroups found, evidencing a clear differentiation from the reference strains. To our knowledge, this is the most extensive study completed for IPNV in Chile, covering isolates from sea- and freshwater salmon farms and showing a high prevalence of this virus in the country.

摘要

传染性胰腺坏死病毒(IPNV)是一种高度传染性疾病的病原体,该疾病在智利鲑鱼养殖中呈地方性流行,给该行业造成了巨大经济损失。在此,我们比较了不同的诊断方法以检测野外样本中的IPNV,包括3种实时逆转录PCR(qRT-PCR)检测、细胞培养分离和间接荧光抗体试验(IFAT)。此外,我们进行了系统发育分析,以研究智利流行的基因群及其地理分布和毒力。所使用的3种qRT-PCR检测方法的引物靶向该病毒的VP2和VP1基因区域,并在46个样本中进行了测试,其结果之间具有较好的一致性。所有样本至少在2种qRT-PCR检测中呈阳性,29个样本在细胞培养中呈阳性,23个样本在IFAT中呈阳性,后两种方法显示出较低的敏感性。对于系统发育分析,通过RT-PCR扩增了A节段VP2区域的1180和523 bp部分,进行测序并与参考菌株以及智利先前研究报告的分离株的序列进行比较。大多数测序分离株属于基因群5(欧洲起源),5个被归类于基因群1(美洲起源)。智利分离株在每个发现的基因群内形成簇,表明与参考菌株有明显差异。据我们所知,这是在智利针对IPNV完成的最广泛的研究,涵盖了海水和淡水鲑鱼养殖场的分离株,并显示该病毒在该国具有很高的流行率。

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