de Smit A J, Eblé P L, de Kluijver E P, Bloemraad M, Bouma A
Department of Mammalian Virology, Institute for Animal Science and Health (ID-DLO), Lelystad, The Netherlands.
Prev Vet Med. 1999 Dec 1;42(3-4):185-99. doi: 10.1016/s0167-5877(99)00075-6.
The National Reference Laboratory for classical swine fever (CSF) virus in The Netherlands examined more than two million samples for CSF virus or serum antibody during the CSF epizootic of 1997-1998. The immense amount of samples and the prevalence of border disease (BD) virus and bovine viral diarrhoea (BVD) virus infections in Dutch pig herds necessitated the diagnostic efforts of the laboratory to be focused on generating CSF specific test results throughout the eradication campaign. Detection of 82% of the 429 outbreaks was achieved through the combined use of a direct immunofluorescence and peroxidase assay (FAT/IPA) with samples (tonsils) collected from clinically-suspected pigs. This suggests that in the majority of the outbreaks, the pigs had clinical signs that were recognised by the farmer and/or veterinarians, indicating the presence of CSF virus in a pig herd. A positive diagnosis of 74% of all the tissue samples (tonsils) collected at infected pig holdings was established by FAT. More than 140,000 heparinised blood samples were examined by virus isolation, resulting in the detection of 4.5% of the infected herds. CSF virus was isolated in approximately 29% of all the blood samples collected from pigs at infected or suspected farms. Several serological surveys--each done within a different framework--led to the detection of 13.5% of the total number of outbreaks. The detection of CSF virus antibody in serum was carried out by semi-automated blocking ELISA. Approximately 28.5% of the sera which reacted in the ELISA were classified as CSF virus-neutralising antibody positive and 26.5% as positive for other pestiviruses following the virus neutralisation test (VNT). We concluded that two of the CSF laboratory diagnostic methods described were determinative in the eradication campaign: first, the FAT for the screening of diseased pigs; and second, the ELISA and VNT when millions of predominantly healthy pigs needed to be screened for the presence of CSF serum antibody. Decision-making on the basis of results generated by either method can, however, be seriously hindered when samples are examined from pig herds with a high prevalence of non-CSF pestiviruses.
荷兰国家猪瘟(CSF)病毒参考实验室在1997 - 1998年猪瘟流行期间检测了超过两百万份样本,以检测猪瘟病毒或血清抗体。由于样本数量巨大,且荷兰猪群中存在边境病(BD)病毒和牛病毒性腹泻(BVD)病毒感染,该实验室的诊断工作在整个根除行动中都集中于生成猪瘟特异性检测结果。通过将直接免疫荧光和过氧化物酶检测(FAT/IPA)与从临床疑似猪采集的样本(扁桃体)联合使用,检测到了429起疫情中的82%。这表明在大多数疫情中,猪出现了被养殖户和/或兽医识别出的临床症状,表明猪群中存在猪瘟病毒。通过FAT对在感染猪场采集的所有组织样本(扁桃体)中的74%做出了阳性诊断。通过病毒分离检测了超过140,000份肝素化血样,结果在4.5%的感染猪群中检测到了猪瘟病毒。在所有从感染或疑似猪场采集的猪的血样中,约29%分离出了猪瘟病毒。几项血清学调查——每项调查都在不同框架下进行——导致检测到了疫情总数的13.5%。血清中猪瘟病毒抗体的检测通过半自动阻断ELISA进行。在ELISA中发生反应的血清中,约28.5%在病毒中和试验(VNT)后被分类为猪瘟病毒中和抗体阳性,26.5%对其他瘟病毒呈阳性。我们得出结论,所描述的两种猪瘟实验室诊断方法在根除行动中具有决定性作用:第一,FAT用于筛查患病猪;第二,ELISA和VNT用于对数以百万计的主要健康猪进行猪瘟血清抗体检测。然而,当从非猪瘟瘟病毒高流行率的猪群中采集样本进行检测时,基于这两种方法产生的结果进行决策可能会受到严重阻碍。