Husson M O, Trinel P A, Izard D, Mielcarek C, Gavini F, Leclerc H
Ann Inst Pasteur Microbiol. 1987 Jan-Feb;138(1):39-48. doi: 10.1016/0769-2609(87)90052-4.
Monoclonal antibodies (MoAb) to the alkaline phosphatase of Escherichia coli were produced from spleen cells of BALB/c mice primed with purified alkaline phosphatase of E. coli and SP2O/Ag-14 myeloma cells. Five stable clones were established. They all produced antibodies which reacted by enzyme-linked immunosorbent assay (ELISA) with alkaline phosphatase of all E. coli (25 strains) independently of their origin (drinking water, saline water, surface water, faecal or clinical origin), and with that of four Shigella species (7 strains) tested. Four of these MoAb gave a positive reaction with 52% (MoAb 4G10), 73% (MoAb 4F8, MoAb 4G6) and 89% (MoAb 3C8) of 14 other bacterial species (30 strains) studied, while one (MoAb 2E5) did not react with alkaline phosphatase of these unrelated bacterial strains and thus appeared specific for E. coli and Shigella species. This MoAb was still detectable in ascitic fluids at 1/500,000 in ELISA, and detected all E. coli strains in an indirect immunofluorescence assay at 1/100. It could therefore be used as a reagent for routine detection of E. coli in drinking water, foods or clinical specimens.
用纯化的大肠杆菌碱性磷酸酶免疫BALB/c小鼠，取其脾细胞与SP2O/Ag - 14骨髓瘤细胞融合，制备出针对大肠杆菌碱性磷酸酶的单克隆抗体（MoAb）。建立了5个稳定的克隆株。通过酶联免疫吸附测定（ELISA）发现，它们所产生的抗体能与所有大肠杆菌（25株）的碱性磷酸酶发生反应，而不论这些大肠杆菌的来源（饮用水、盐水、地表水、粪便或临床来源），并且能与所检测的4种志贺氏菌属（7株）的碱性磷酸酶发生反应。其中4种单克隆抗体与所研究的其他14种细菌（30株）中的52%（单克隆抗体4G10）、73%（单克隆抗体4F8、单克隆抗体4G6）和89%（单克隆抗体3C8）的碱性磷酸酶呈阳性反应，而有一种（单克隆抗体2E5）不与这些不相关细菌菌株的碱性磷酸酶发生反应，因此似乎对大肠杆菌和志贺氏菌属具有特异性。在ELISA中，该单克隆抗体在腹水液中的检测浓度可达1/500,000，在间接免疫荧光测定中以1/100的浓度可检测到所有大肠杆菌菌株。因此，它可用作饮用水、食品或临床标本中大肠杆菌常规检测的试剂。
