一种快捷、定量检测烟曲霉GM抗原双mAb夹心ELISA的方法

Development of a rapid quantitative double mAb sandwich ELISA for detecting galactomannan antigen of aspergillus fumigatus

目的 :建立一种快速诊断烟曲霉病的双mAb夹心ELISA法。方法 :应用 4株抗烟曲霉GM单克隆抗体 (mAb) ,分别包被和制备HRP mAb ,用双mAb夹心ELISA法配对试验 ,选择捕获及检测mAb。结果 :经筛选得到捕获及检测mAb的最佳组合 ,并建立了双mAb夹心ELISA法。该法检测GM抗原的灵敏度为 0 .1μg/L ,测出范围在 0 .1～ 10 μg/L之间。连续 6d用ELISA法检测同一份样品 ,所获CV的平均值为 ( 7.2± 3.8) %。结论 :建立了一种可快速、定量检测烟曲霉GM抗原的双mAb夹心ELISA法 ,灵敏度高、重复性好 ,对研制试剂盒应用于烟曲霉病早期诊断和防治 。

AIM: To develop a double mAb sandwich ELISA for rapidly and quantitatively detecting galactomannan (GM) antigen of aspergillus fumigatus. METHODS: Four monoclonal antibodies (mAb) against GM of aspergillus fumigatus were used as coating or enzyme conjugated antibodies respectively. Capture and sandwich mAbs were selected by sandwich ELISA paired one by one. RESULTS: Optimal capture and sandwich mAbs were selected and a highly sensitive double sandwich ELISA established. The sensitivity of detecting GM reached 0.1 μg/L, the detectable range was 0.1～10 μg/L . Coefficient of variation (C.V) obtained from detecting the same sample for 6 times by ELISA was (7.2±3.8)% . CONCLUSION: A sensitive, repeatable and rapid double mAb sandwich ELISA was established for quantitation of aspergillus fumigatus GM, which might apply to early diagnosis and treatment of patients with aspergillosis.