Wu S J, Pacheco N D, Oprandy J J, Rollwagen F M
Infectious Diseases Department, Naval Medical Research Institute, Bethesda, Maryland 20889.
Infect Immun. 1991 Aug;59(8):2555-9. doi: 10.1128/iai.59.8.2555-2559.1991.
The development of a rapid and specific diagnostic assay for Campylobacter infections is important in determining the etiology of acute diarrhea in humans. Studies have shown that sonicated whole bacteria or partially purified antigens cross-reacted with antibodies against other closely related bacteria. To solve the problems of specificity, we identified specific antigens of Campylobacter jejuni and Campylobacter coli for use in diagnostic assays. We investigated the responses of serum, urine, and intestinal lavage antibodies in infected (fed live bacteria) and parenterally immunized (intraperitoneal injection of sonicated whole bacteria with adjuvant) mice directed against C. jejuni or C. coli by Western blot (immunoblot) analysis. Antibody responses were examined weekly for up to 28 days. Fewer antigens were detected by urinary and intestinal lavage fluid immunoglobulin A (IgA) than serum IgG and IgM for both parenterally immunized and infected mice. Serum from parenterally immunized mice detected more antigens than that from infected mice. Two high-molecular-weight antigens (62,000 and 43,000) were predominantly detected by serum, urine, and intestinal lavage fluids of both parenterally immunized and infected mice. Serum antibodies from 28-day parenterally immunized mice detected one antigen specific to C. coli with a molecular weight of 38,000 and one antigen specific to C. jejuni with a molecular weight of 27,000. An immunodominant protein with a molecular weight of 31,000 common to both C. jejuni and C. coli was also recognized by serum antibodies from parenterally immunized mice.
开发一种针对弯曲杆菌感染的快速、特异性诊断检测方法对于确定人类急性腹泻的病因很重要。研究表明，超声破碎的全菌或部分纯化的抗原会与针对其他密切相关细菌的抗体发生交叉反应。为了解决特异性问题，我们鉴定了空肠弯曲杆菌和结肠弯曲杆菌的特异性抗原，用于诊断检测。我们通过蛋白质印迹（免疫印迹）分析，研究了感染（喂食活细菌）和经肠外免疫（腹腔注射超声破碎的全菌与佐剂）小鼠的血清、尿液和肠道灌洗液中针对空肠弯曲杆菌或结肠弯曲杆菌的抗体反应。每周检查抗体反应，最长持续28天。对于经肠外免疫和感染的小鼠，尿液和肠道灌洗液中的免疫球蛋白A（IgA）检测到的抗原比血清IgG和IgM少。经肠外免疫小鼠的血清检测到的抗原比感染小鼠的多。经肠外免疫和感染的小鼠的血清、尿液和肠道灌洗液主要检测到两种高分子量抗原（62,000和43,000）。经28天肠外免疫小鼠的血清抗体检测到一种分子量为38,000的结肠弯曲杆菌特异性抗原和一种分子量为27,000的空肠弯曲杆菌特异性抗原。经肠外免疫小鼠的血清抗体还识别出一种空肠弯曲杆菌和结肠弯曲杆菌共有的分子量为31,000的免疫显性蛋白。
