Notermans S, Wernars K
Laboratory of Water and Food Microbiology, National Institute of Public Health and Environmental Hygiene, Bilthoven, The Netherlands.
Int J Food Microbiol. 1990 Aug;11(1):35-49. doi: 10.1016/0168-1605(90)90038-7.
During the last decade several new analytical techniques have been developed for testing food products and clinical samples. One technique uses sensitive immunoassays such as enzyme-linked immunosorbent assay (ELISA) and latex agglutination. The most important step in developing sensitive immunoassays is the evaluation of the assay for specificity, cross-reactivity and sensitivity. False-negative results can easily be detected by adding known quantities of antigen to the sample. The most appropriate way to detect false-positive results is the specific inhibition of the immunological reaction by addition to the test-sample of either synthetic epitopes or anti-idiotype antibodies. The progress in recombinant DNA techniques now offers opportunities for application as analytical tools in food and clinical microbiology. Methods are being developed to detect microorganisms by their nucleic acid sequence using the so-called hybridization procedure. With this technique, labelled DNA fragments (probes) are hybridized with a complementary base sequence present in the microorganism. Foodborne pathogens can be detected by using a probe with a complementary base sequence which codes for toxin production. DNA-DNA hybridization techniques may replace the traditional cultural techniques for assaying pathogenic micro-organisms. However, more experience with these techniques is needed before further evaluation can be given.
在过去十年间,已开发出多种用于检测食品和临床样本的新分析技术。其中一种技术采用诸如酶联免疫吸附测定(ELISA)和乳胶凝集等灵敏的免疫测定法。开发灵敏免疫测定法的最重要步骤是评估该测定法的特异性、交叉反应性和灵敏度。通过向样本中添加已知量的抗原,可轻松检测到假阴性结果。检测假阳性结果的最合适方法是向测试样本中添加合成表位或抗独特型抗体,特异性抑制免疫反应。重组DNA技术的进展为其作为食品和临床微生物学中的分析工具提供了应用机会。目前正在开发利用所谓杂交程序通过微生物的核酸序列检测微生物的方法。利用该技术,标记的DNA片段(探针)与微生物中存在的互补碱基序列杂交。通过使用具有编码毒素产生的互补碱基序列的探针,可检测食源性病原体。DNA-DNA杂交技术可能会取代用于检测病原微生物的传统培养技术。然而,在能够进行进一步评估之前,还需要积累更多使用这些技术的经验。