Novel Approach for Isolation and Identification of Porcine Epidemic Diarrhea Virus (PEDV) Strain NJ Using Porcine Intestinal Epithelial Cells.
Novel Approach for Isolation and Identification of Porcine Epidemic Diarrhea Virus (PEDV) Strain NJ Using Porcine Intestinal Epithelial Cells.
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使用猪肠上皮细胞分离和鉴定猪流行性腹泻病毒 (PEDV) NJ 株的新方法
DOI：
10.3390/v9010019
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发表时间：
2017-01-21
期刊：
影响因子：
--
通讯作者：
Li Y
中科院分区：
文献类型：
--
作者：
Shi W;Jia S;Zhao H;Yin J;Wang X;Yu M;Ma S;Wu Y;Chen Y;Fan W;Xu Y;Li Y
猪流行性腹泻病毒（Porcine epidemic diarrhea virus，PEDV）是引起国内外猪流行性腹泻病的病原，给养猪业造成了严重的经济损失。PEDV灭活疫苗是控制PEDV流行的关键。然而，在PEDV体外繁殖期间获得一致的低病毒滴度;这代表了对病毒和疫苗开发的分子分析的挑战。在这项研究中，我们成功地分离了PEDV分离株（NJ株），从临床样品中收集在最近爆发的仔猪腹泻在中国，使用猪肠上皮细胞（IEC）。我们发现，分离物更好地适应IEC比在Vero细胞中的生长，IEC培养物的滴度为104.5 TCID 50/0.1 mL，在第45代。随着病毒传代，S蛋白的突变增加，并且突变倾向于减毒。病毒攻击表明，IEC适应培养物在第45代的存活率高于第5代。利用IEC分离和增殖PEDV为PEDV的实验室诊断、流行病学特征和分子生物学研究提供了一种有效的方法。
Porcine epidemic diarrhea virus (PEDV), which is the causative agent of porcine epidemic diarrhea in China and other countries, is responsible for serious economic losses in the pork industry. Inactivated PEDV vaccine plays a key role in controlling the prevalence of PEDV. However, consistently low viral titers are obtained during the propagation of PEDV in vitro; this represents a challenge to molecular analyses of the virus and vaccine development. In this study, we successfully isolated a PEDV isolate (strain NJ) from clinical samples collected during a recent outbreak of diarrhea in piglets in China, using porcine intestinal epithelial cells (IEC). We found that the isolate was better adapted to growth in IECs than in Vero cells, and the titer of the IEC cultures was 104.5 TCID50/0.1 mL at passage 45. Mutations in the S protein increased with the viral passage and the mutations tended towards attenuation. Viral challenge showed that the survival of IEC-adapted cultures was higher at the 45th passage than at the 5th passage. The use of IECs to isolate and propagate PEDV provides an effective approach for laboratory-based diagnosis of PEDV, as well as studies of the epidemiological characteristics and molecular biology of this virus.
影响因子：
5
作者：
Correa I;Jiménez G;Suñé C;Bullido MJ;Enjuanes L
通讯作者：
Enjuanes L