Rapid and highly sensitive portable detection of African swine fever virus
Rapid and highly sensitive portable detection of African swine fever virus
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DOI：
10.1111/tbed.13770
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发表时间：
2020-08-31
影响因子：
4.3
通讯作者：
Ambagala, Aruna
中科院分区：
文献类型：
--
作者：
Daigle, Jade;Onyilagha, Chukwunonso;Ambagala, Aruna
非洲猪瘟（ASF）继续在亚洲蔓延，毁灭了猪群。该病在猪中几乎是100%致命的，目前还没有有效的疫苗。因此，早期发现ASF对于有效控制疾病至关重要。检测过程通常需要将样本运送到中心实验室，这可能需要数小时的旅行或运输时间，从而延迟快速响应所需的结果。在现场或技术能力和/或基础设施有限的区域实验室确认ASFV感染动物的能力应消除这些问题。本研究描述了将高度灵敏度和特异性的实验室验证的实时PCR检测成功转移到便携式笔侧热循环仪上，该热循环仪可以在现场操作，在快速手动核酸提取后快速检测ASFV从广泛的临床样本中，包括口腔液等聚集体样本。便携式检测试剂盒的性能与基于实验室的检测试剂盒相当。在越南的7个疑似ASF农场，通过现场检测89个新鲜采集的全血样本，评估了该检测试剂盒的真正便携性。现场获得的结果与第二天获得的实验室数据一致。在需要快速实验室结果时，这种可现场部署的分子测定的可用性将消除将样本运送到中心实验室的需要，最终改善响应时间。
African swine fever (ASF) continues to spread across Asia, devastating pig populations. The disease is nearly 100% fatal in pigs, and currently, there is no effective vaccine available. Therefore, early detection of ASF is critical for effective disease control. The testing process usually requires samples to be shipped to a central laboratory, which may take many hours of travel or shipping time, delaying the results needed for a rapid response. The ability to confirm ASFV-infected animals on-site or in a regional laboratory that has limited technical capacity and/or infrastructure should eliminate these issues. This study describes the successful transfer of a highly sensitive and specific laboratory-validated real-time PCR assay to a portable pen-side thermocycler, which can be operated in the field for rapid detection of ASFV following a quick manual nucleic acid extraction from a wide array of clinical samples including aggregate samples such as oral fluids. The performance of the portable assay was comparable to the laboratory-based assay. The true portability of the assay was evaluated in seven ASF-suspected farms in Vietnam by testing eighty-nine freshly collected whole blood samples on-site. The results obtained on-site were in agreement with the laboratory data obtained the following day. Availability of this field-deployable molecular assay would eliminate the need to ship samples to a central laboratory, when rapid laboratory results are required, ultimately improving the response time.