McCullough K C
Institut für Viruskrankheiten und Immunprophylaxe, Mittelhäusern, Switzerland.
Rev Sci Tech. 1993 Jun;12(2):325-53. doi: 10.20506/rst.12.2.696.
Methodologies for animal disease diagnosis and the analysis and control of animal health currently make much use of immunoassay techniques, in particular ELISA, and tests based on nucleic acid measurement (nucleic acid hybridisation, polymerase chain reaction [PCR]). In the latter area, the recently developed PCRs have become most useful. Recombinant technologies have yielded defined and extremely pure antigens, which can give more accurate interpretations of results obtained using diagnostic tests; in addition, such antigens obviate the need to handle live pathogens, except where these may be contained within the sample for diagnosis. The application of monoclonal antibodies (MAbs) to many tests, in particular those which use immunoassay bases or perform molecular fingerprinting of isolates, has increased the specificity and usefulness of these methodologies. Modern technology has permitted a considerable reduction in the use of animals for MAb generation through in vitro immunisation methods and the application of serum-free media with bio-reactors. These in vitro immunisation procedures have an additional application in the analysis of vaccine efficacy without recourse to animal experimentation. Furthermore, in vitro methods can be used to monitor the immune response in an animal and, in particular, to measure the development of immunological memory. It is now possible to estimate more accurately the quality and duration of an induced immune response without using animals for testing. Not only are the assay systems developed through biotechnology more sensitive and less hazardous than previous techniques (especially with regard to the necessity of handling live pathogens), but they can also determine more accurately the immune responsiveness or immune status of the animals under study.
目前,动物疾病诊断以及动物健康分析与控制的方法大量使用免疫测定技术,尤其是酶联免疫吸附测定(ELISA),以及基于核酸测量的检测方法(核酸杂交、聚合酶链反应[PCR])。在后一领域,最近开发的PCR技术已变得极为有用。重组技术已产生了明确且纯度极高的抗原,这些抗原能够对诊断检测所获得的结果给出更准确的解释;此外,除了样本中可能含有用于诊断的活病原体之外,此类抗原无需处理活病原体。单克隆抗体(MAb)在许多检测中的应用,特别是那些基于免疫测定或对分离株进行分子指纹识别的检测,提高了这些方法的特异性和实用性。现代技术通过体外免疫方法以及在生物反应器中使用无血清培养基,使得用于产生单克隆抗体的动物数量大幅减少。这些体外免疫程序在不借助动物实验的情况下,在疫苗效力分析中还有额外的应用。此外,体外方法可用于监测动物的免疫反应,特别是用于测量免疫记忆的发展。现在有可能在不使用动物进行检测的情况下,更准确地估计诱导免疫反应的质量和持续时间。通过生物技术开发的检测系统不仅比以前的技术更灵敏、危害更小(特别是在处理活病原体的必要性方面),而且还能更准确地确定所研究动物的免疫反应性或免疫状态。