Department of Virology, Central Veterinary Institute of Wageningen UR, P.O. Box 65, 8200 AB Lelystad, The Netherlands.
Vet Microbiol. 2010 Mar 24;141(3-4):275-81. doi: 10.1016/j.vetmic.2009.09.028. Epub 2009 Sep 26.
A considerable part of tissue samples that are collected for the monitoring of classical swine fever (CSF) from the wild boar population or from domestic pigs are unsuitable for virus detection using the fluorescent antibody test (FAT) or virus isolation (VI), due to tissue degradation. Reverse transcription polymerase chain reaction (RT-PCR) has a higher sensitivity than the FAT and VI, and is supposed to be less sensitive to sample degradation. Reliable and quantitative information on how long viral RNA and infectious virus can be detected in organs and which organs are most susceptible to degradation is, however, lacking. In the present study we generated survival curves of infectious CSF virus (CSFV) and viral RNA in the tonsil, mesenteric lymph node, spleen and kidney, obtained from 24 pigs infected with a moderately virulent CSFV strain. Tissue samples were stored at room temperature and tested by VI and RT-PCR, directly after storage and 1, 2, 3, 4, 7, 14 and 21 days later. It was shown that the RT-PCR is not only more sensitive than VI on fresh tissue samples, but RT-PCR is also less vulnerable to sample degradation. Average half-life values of viral RNA in the tissues ranged from 0.95 to 2.55 days, while half-life values of infectious virus ranged from 0.21 to 0.31 days. The tonsil and spleen are regarded as the most appropriate organs for the detection of infectious virus and viral RNA, not only in fresh samples, but also in samples that suffer from tissue degradation.
相当一部分用于监测野猪或家猪群体中经典猪瘟（CSF）的组织样本，由于组织降解，不适合使用荧光抗体试验（FAT）或病毒分离（VI）进行病毒检测。逆转录聚合酶链反应（RT-PCR）比 FAT 和 VI 具有更高的灵敏度，并且据推测对样品降解的敏感性较低。然而，关于在器官中可以检测到病毒 RNA 和传染性病毒的时间以及哪些器官最容易降解，缺乏可靠和定量的信息。在本研究中，我们从感染中等毒力 CSFV 株的 24 头猪中获得了扁桃体、肠系膜淋巴结、脾脏和肾脏中的传染性 CSF 病毒（CSFV）和病毒 RNA 的存活曲线。组织样本在室温下储存，并通过 VI 和 RT-PCR 直接在储存后以及 1、2、3、4、7、14 和 21 天后进行检测。结果表明，RT-PCR 不仅在新鲜组织样本上比 VI 更敏感，而且 RT-PCR 对样品降解的敏感性也较低。组织中病毒 RNA 的平均半衰期值范围为 0.95 至 2.55 天，而传染性病毒的半衰期值范围为 0.21 至 0.31 天。扁桃体和脾脏被认为是检测传染性病毒和病毒 RNA 的最适宜器官，不仅在新鲜样本中，而且在组织降解的样本中也是如此。
