Pre-Clinical Evaluation of a Real-Time PCR Assay on a Portable Instrument as a Possible Field Diagnostic Tool: Experiences from the Testing of Clinical Samples for African and Classical Swine Fever Viruses
Pre-Clinical Evaluation of a Real-Time PCR Assay on a Portable Instrument as a Possible Field Diagnostic Tool: Experiences from the Testing of Clinical Samples for African and Classical Swine Fever Viruses
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DOI：
10.1111/tbed.12538
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发表时间：
2017-10-01
影响因子：
4.3
通讯作者：
Belak, S.
中科院分区：
文献类型：
--
作者：
Liu, L.;Luo, Y.;Belak, S.
非洲猪瘟（ASF）和经典猪瘟（CSF）是两种高度传染性的跨界动物疾病（TADs），对全球养猪业构成严重威胁，包括世界上最大的猪肉生产国中国。本研究建立了一种快速鉴别非洲猪瘟病毒（ASFV）和猪瘟病毒（CSFV）的双重荧光定量PCR方法。该测定在便携式电池供电的PCR热循环仪上进行，具有低样品通量（称为“T-COR 4测定”）。通过检测在中国采集的临床样本，研究了T-COR 4测定作为可能的现场方法的可行性和可靠性。当用参考材料或实验感染样品进行评价时，该检测方法以可靠的方式进行，产生的结果与从固定PCR平台获得的结果相当。在59份临床样本中，41份的结果与两步CSFV实时PCR检测结果相同。在这些样品中未检测到ASFV。T-COR 4测定在技术上易于进行，并在3小时内产生结果，包括样品制备。结合简单的样品制备方法，T-COR 4检测为ASF和CSF的现场诊断和鉴别提供了新的工具，这在偏远地区可能具有特别的价值。
African swine fever (ASF) and classical swine fever (CSF) are two highly infectious transboundary animal diseases (TADs) that are serious threats to the pig industry worldwide, including in China, the world's largest pork producer. In this study, a duplex real-time PCR assay was developed for the rapid detection and differentiation of African swine fever virus (ASFV) and classical swine fever virus (CSFV). The assay was performed on a portable, battery-powered PCR thermocycler with a low sample throughput (termed as T-COR4 assay'). The feasibility and reliability of the T-COR4 assay as a possible field method was investigated by testing clinical samples collected in China. When evaluated with reference materials or samples from experimental infections, the assay performed in a reliable manner, producing results comparable to those obtained from stationary PCR platforms. Of 59 clinical samples, 41 had results identical to a two-step CSFV real-time PCR assay. No ASFV was detected in these samples. The T-COR4 assay was technically easy to perform and produced results within 3h, including sample preparation. In combination with a simple sample preparation method, the T-COR4 assay provides a new tool for the field diagnosis and differentiation of ASF and CSF, which could be of particular value in remote areas.