Domingues Helena Gallicchio, Campalans Jacqueline, Almeida Renata Servan, Coswig Lia Treptow, Arns Clarice Weis
Departamento de Microbiologia e Imunologia, Instituto de Biologia, Universidade Estadual de Campinas-UNICAMP, SP, Brazil.
Vet Res. 2002 Jul-Aug;33(4):397-404. doi: 10.1051/vetres:2002025.
A dot-ELISA test for the detection of anti-BRSV antibodies is described. The objective of this study was the standardisation of a test as a fast, inexpensive and effective alternative to detect anti-BRSV antibodies. Its sensitivity, specificity and usefulness were compared to a commercial ELISA-kit and to the standard serum neutralisation (SN) test. The standardisation of the technique was done using nitrocellulose disks soaked with a viral sample isolated in Brazil, BRSV-25-BR. The best results were obtained when the disks were sensitised with a purified antigen at a concentration of 0.7 microg/disk and the bovine serum was diluted 1: 200. The experiment used 423 samples of bovine serum collected in the main cattle breeding centres in Brazil. The standard SN, dot-ELISA technique and commercial ELISA kits scored 67.8%, 71.8% and 72.3% of the samples as positive, respectively. When compared to the SN test, the standardised dot-ELISA and the commercial ELISA tests presented relative sensitivities of 92.3% and 91.6% and relative specificities of 71.3% and 68.4% respectively. The results demonstrated that the dot-ELISA test is adequate for the objectives proposed by this study, being easy to use and economically viable. Thus, this test represents an alternative for BRSV serological diagnosis in the substitution of SN and commercial ELISA tests, recommendable for utilisation in laboratories with few resources.
本文描述了一种用于检测抗牛呼吸道合胞病毒(BRSV)抗体的斑点酶联免疫吸附测定(dot-ELISA)试验。本研究的目的是将该试验标准化,作为一种快速、廉价且有效的检测抗BRSV抗体的替代方法。将其敏感性、特异性和实用性与一种商业酶联免疫吸附测定试剂盒以及标准血清中和(SN)试验进行了比较。该技术的标准化是使用用在巴西分离的病毒样本BRSV-25-BR浸泡的硝酸纤维素圆盘完成的。当圆盘用浓度为0.7微克/圆盘的纯化抗原致敏且牛血清稀释1:200时,获得了最佳结果。该实验使用了在巴西主要养牛中心收集的423份牛血清样本。标准SN试验、斑点酶联免疫吸附测定技术和商业酶联免疫吸附测定试剂盒分别将67.8%、71.8%和72.3%的样本判定为阳性。与SN试验相比,标准化的斑点酶联免疫吸附测定和商业酶联免疫吸附测定试验的相对敏感性分别为92.3%和91.6%,相对特异性分别为71.3%和68.4%。结果表明,斑点酶联免疫吸附测定试验符合本研究提出的目标,易于使用且经济可行。因此,该试验是替代SN试验和商业酶联免疫吸附测定试验用于BRSV血清学诊断的一种选择,推荐在资源有限的实验室使用。