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Evaluation of FTA (R) card for the rescue of infectious foot-and-mouth disease virus by chemical transfection of extracted RNA in cultured cells

机译:通过化学转染培养细胞中的RNA拯救FTA(R)卡以挽救传染性口蹄疫病毒的评估

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Foot-and-mouth disease (FMD) is a highly contagious epidemic disease of transboundary importance. Inadequate storage and shipment of suspected clinical samples can compromise the ability to detect and characterise FMD virus (FMDV) in endemic countries, thereby, leading to the loss of valuable virological and epidemiological data. This study, investigates the potential of using FTA (R) cards for dry transportation of clinical samples and subsequent recovery of infectious FMDV by chemical transfection of FTA (R) card fixed RNA as an alternative to the conventional cell culture based virus isolation method. A higher proportion of infectious FMDV was rescued from clinical samples (cell culture isolates, tongue epithelial suspension and impression smears) by the FTA (R) card fixed RNA transfection method (76%) compared to the conventional cell culture based virus isolation (56%), suggesting a better performance of the current RNA transfection procedure. Furthermore, it was possible to rescue live virus by the transfection of RNA extracted from FTA (R) card impregnated with clinical samples that had been stored at varying temperature (4-37 degrees C) up to a period of six weeks. The VP1 sequence data and antigenic relationships with the vaccine strains, between viruses rescued by FTA (R) card fixed RNA transfection and conventional cell culture, were comparable. Therefore, these results support the use of the FTA (R) card for the economic, dry, non-hazardous transport of FMD suspected clinical samples from the site of collection to national/international reference laboratories. (C) 2016 Elsevier Ltd. All rights reserved.
机译:口蹄疫(FMD)是具有跨界重要性的高度传染性流行病。可疑临床样品的存储和运输不充分会损害在流行国家检测和鉴定口蹄疫病毒(FMDV)的能力,从而导致有价值的病毒学和流行病学数据丢失。这项研究调查了使用FTA(R)卡进行临床样品的干运输以及随后通过化学转染FTA(R)卡固定RNA作为传统基于细胞培养的病毒分离方法的替代方法恢复传染性FMDV的潜力。与传统的基于细胞培养的病毒分离方法(56%)相比,通过FTA(R)卡固定RNA转染方法从临床样品(细胞培养分离物,舌上皮悬浮液和印模涂片)中拯救了更高比例的传染性FMDV(76%) ),表明目前的RNA转染程序具有更好的性能。此外,有可能通过转染从FTA(R)卡提取的RNA来挽救活病毒,该RNA浸有已在不同温度(4-37摄氏度)下保存长达六周的临床样品。通过FTA(R)卡固定RNA转染挽救的病毒与常规细胞培养物之间的VP1序列数据和与疫苗株的抗原关系具有可比性。因此,这些结果支持使用FTA(R)卡将FMD可疑临床样品从收集地点经济,干燥,无害地运输到国家/国际参考实验室。 (C)2016 Elsevier Ltd.保留所有权利。

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