Engstrom S M, Shoop E, Johnson R C
Department of Microbiology, University of Minnesota Medical School, Minneapolis.
J Clin Microbiol. 1995 Feb;33(2):419-27. doi: 10.1128/jcm.33.2.419-427.1995.
We monitored the antibody responses of 55 treated patients with early Lyme disease and physician-documented erythema migrans. Six sequential serum samples were obtained from patients before, during, and until one year after antibiotic therapy and analyzed by in-house enzyme-linked immunosorbent (ELISA) and immunoblot assays. An immunoblot procedure utilizing a gradient gel and an image analysis system was developed. A relational database management system was used to analyze the results and provide criteria for early disease immunoblot interpretation. Recommended criteria for the immunoglobulin M (IgM) immunoblot are the recognition of two of three proteins (24, 39, and 41 kDa). The recommended criteria for a positive IgG immunoblot are the recognition of two of five proteins (20, 24 [> 19 intensity units], 35, 39, and 88 kDa). Alternatively, if band intensity cannot be measured, the 22-kDa protein can be substituted for the 24-kDa protein with only a small decrease in sensitivity. Monoclonal antibodies were used to identify all these proteins except the 35-kDa protein. With the proposed immunoblot interpretations, the sequential serum samples were examined. At visit 1, the day of diagnosis and initiation of treatment, 54.5% of the serum samples were either IgM or IgG positive. The peak antibody response, with 80% of the serum samples positive, occurred at visit 2, 8 to 12 days into treatment. The sensitivities of the IgM and IgG immunoblot for detecting patients that were seropositive into the study period were 58.5 and 54.6%, respectively, at visit 1 and 100% at visit 2. Twenty percent of the patients remained seronegative throughout the study. The specificities of the IgM and IgG immunoblots were 92 to 94% and 93 to 96%, respectively. The IgM immunoblot and ELISA were similar in sensitivities, whereas the IgG immunoblot had greater sensitivity than the IgG ELISA (P = 0.006).
我们监测了55例经治疗的早期莱姆病患者和有医生记录的游走性红斑患者的抗体反应。在抗生素治疗前、治疗期间以及治疗后一年，从患者处采集了六份连续的血清样本，并通过内部酶联免疫吸附测定（ELISA）和免疫印迹分析进行检测。开发了一种利用梯度凝胶和图像分析系统的免疫印迹程序。使用关系数据库管理系统分析结果，并为早期疾病免疫印迹解释提供标准。免疫球蛋白M（IgM）免疫印迹的推荐标准是识别三种蛋白质（24、39和41 kDa）中的两种。IgG免疫印迹阳性的推荐标准是识别五种蛋白质（20、24[强度单位>19]、35、39和88 kDa）中的两种。或者，如果无法测量条带强度，则可以用22 kDa蛋白质替代24 kDa蛋白质，灵敏度仅略有下降。除35 kDa蛋白质外，所有这些蛋白质均使用单克隆抗体进行鉴定。根据提出的免疫印迹解释，对连续的血清样本进行了检查。在第1次就诊时，即诊断和开始治疗的当天，54.5%的血清样本IgM或IgG呈阳性。抗体反应峰值出现在第2次就诊时，即治疗开始8至12天，80%的血清样本呈阳性。在第1次就诊时，IgM和IgG免疫印迹检测研究期间血清阳性患者的灵敏度分别为58.5%和54.6%，在第2次就诊时为100%。20%的患者在整个研究期间血清一直呈阴性。IgM和IgG免疫印迹的特异性分别为92%至94%和93%至96%。IgM免疫印迹和ELISA的灵敏度相似，而IgG免疫印迹的灵敏度高于IgG ELISA（P = 0.006）。
