Bühling F, Koch G, Wex T, Heimburg A, Vieth M, Leodolter A, Roessner A, Ansorge S, Malfertheiner P
Institute of Immunology, Otto-von-Guericke University Magdeburg, Germany.
Clin Diagn Lab Immunol. 2004 Jan;11(1):131-6. doi: 10.1128/cdli.11.1.131-136.2004.
Helicobacter pylori is the key pathogen for gastroduodenal diseases. The clinical outcome of H. pylori infection is influenced by the presence of strain-specific virulence factors that are usually detected by the presence of specific anti-H. pylori antibodies in serum. Apart from the detection of these antibodies by enzyme-linked immunosorbent assay (ELISA), it is desirable to obtain additional information concerning the presence of certain virulence factors of H. pylori that are currently detected by immunoblot analysis. At present, the immunodiagnosis of an H. pylori infection includes two separate methods: ELISA and immunoblot analysis. Here, we report the development and evaluation of a new rapid flow microparticle immunofluorescence assay (FMIA) for detection of anti-H. pylori antibodies in human serum. The assay allows rapid qualitative and quantitative detection of anti-H. pylori antibodies by using crude antigen preparations as well as single recombinant antigens (urease A, urease B, CagA, and alkylhydroxy peroxide reductase) in the same sample with one measurement, and thus it combines the advantages of enzyme immunoassay and Western blot analysis. Seventy-five patient samples were analyzed by FMIA, ELISA, and Western blotting with respect to their immunoreactivity against crude H. pylori extracts and individual H. pylori antigens. Statistical analyses revealed an overall similarity of more than 90% among the results for FMIA, ELISA, and Western blot. Therefore, we conclude that FMIA is a powerful and time- and cost-saving assay system for the detection of antimicrobial antibodies, with higher sensitivity and a larger measurement range than ELISA.
幽门螺杆菌是胃十二指肠疾病的关键病原体。幽门螺杆菌感染的临床结果受菌株特异性毒力因子的影响，这些毒力因子通常通过血清中特异性抗幽门螺杆菌抗体的存在来检测。除了通过酶联免疫吸附测定（ELISA）检测这些抗体外，还希望获得有关目前通过免疫印迹分析检测到的幽门螺杆菌某些毒力因子存在的更多信息。目前，幽门螺杆菌感染的免疫诊断包括两种独立的方法：ELISA和免疫印迹分析。在此，我们报告了一种用于检测人血清中抗幽门螺杆菌抗体的新型快速流动微粒免疫荧光测定（FMIA）的开发和评估。该测定法允许通过在同一样品中使用粗抗原制剂以及单一重组抗原（脲酶A、脲酶B、细胞毒素相关蛋白A和烷基过氧化氢还原酶）进行一次测量，快速定性和定量检测抗幽门螺杆菌抗体，因此它结合了酶免疫测定和蛋白质印迹分析的优点。通过FMIA、ELISA和蛋白质印迹法分析了75份患者样品对幽门螺杆菌粗提取物和单个幽门螺杆菌抗原的免疫反应性。统计分析显示，FMIA、ELISA和蛋白质印迹法的结果总体相似度超过90%。因此，我们得出结论，FMIA是一种用于检测抗菌抗体的强大且节省时间和成本的测定系统，与ELISA相比具有更高的灵敏度和更大的测量范围。
