Zhejiang Medical College, Hangzhou, China.
Biomed Environ Sci. 2011 Jun;24(3):291-9. doi: 10.3967/0895-3988.2011.03.013.
To construct a lipL32/1-lipL21-OmpL1/2 fusion gene and its prokaryotic expression system, and to establish an enzyme-linked immunosorbent assay (ELISA) using the rLipL32/1-LipL21-OmpL1/2 fusion antigen of Leptospira interrogans for sensitive and specific detection of IgM in the serum of patients with leptospirosis.
lipL32/1-lipL21-OmpL1/2 fusion genes were constructed using a primer-linking PCR. The target recombinant protein antigens, rLipL32/1, rLipL21, rOmpL1/2 and rLipL32/1-LipL21-OmpL1/2, were expressed and the purified antigens were then immobilized to the surface of microplate wells for ELISA-based detection of IgM in the sera of leptospirosis patients.
Of 493 acute leptospirosis patients, 95.7% and 97.8% were positive by rLipL32/1-LipL21- OmpL1/2-IgM-ELISA using different serum dilutions, which was higher than the rLipL32/1-IgM-ELISA (93.1% and 90.3%), rLipL21-IgM-ELISA (90.3% and 87.0%), and rOmpL1-IgM-ELISA (85.6% and 81.1%) (P<0.01). All IgM-ELISAs tested negative against 56 non-leptospirosis patients with typhoid fever, hemorrhagic fever or dengue fever.
Trigeminal fusion antigen increases ELISA sensitivity and the rLipL32/1-LipL21-OmpL1/2- IgM-ELISA is a sensitive and specific serological diagnostic method for clinical leptospirosis.
构建脂磷壁酸 L32/1-脂磷壁酸 L21-OmpL1/2 融合基因及其原核表达系统，并利用钩端螺旋体 LipL32/1-LipL21-OmpL1/2 融合抗原建立酶联免疫吸附试验（ELISA），用于灵敏、特异地检测钩端螺旋体病患者血清中的 IgM。
采用引物连接 PCR 构建 lipL32/1-lipL21-OmpL1/2 融合基因。表达目的重组蛋白抗原 rLipL32/1、rLipL21、rOmpL1/2 和 rLipL32/1-LipL21-OmpL1/2，并将纯化的抗原固定在酶联免疫吸附试验（ELISA）板孔表面，用于检测钩端螺旋体病患者血清中的 IgM。
493 例急性钩端螺旋体病患者，rLipL32/1-LipL21-OmpL1/2-IgM-ELISA 采用不同血清稀释度检测，阳性率分别为 95.7%和 97.8%，均高于 rLipL32/1-IgM-ELISA（93.1%和 90.3%）、rLipL21-IgM-ELISA（90.3%和 87.0%）和 rOmpL1-IgM-ELISA（85.6%和 81.1%）（P<0.01）。所有 IgM-ELISA 检测 56 例伤寒、出血热或登革热非钩端螺旋体病患者血清均为阴性。
三价融合抗原可提高 ELISA 检测的敏感性，rLipL32/1-LipL21-OmpL1/2-IgM-ELISA 是一种灵敏、特异的临床钩端螺旋体病血清学诊断方法。
