全血快速检测日本血吸虫抗体免疫层析法的研制和初步评价

Development and Evaluation of a Rapid Immunochromatography Assay for Detection of the Antibody against Schistosoma japonica in Whole-Blood Sample

本研究采用抗人IgG单克隆抗体标记红色乳胶微球作为探针,以日本血吸虫虫卵可溶性抗原包被硝酸纤维素膜,运用免疫间接法原理,建立了全血快速检测血吸虫抗体乳胶免疫层析法（WB-DLIA）.以WB-DLIA和ELISA分别检测粪检血吸虫虫卵阳性病人标本,阳性符合率为94.40%（51/54）和96.30%（52/54）;检测正常人标本,阴性符合率分别为96.6%（252/261）和 95.40%（249/261）,两法之间无显著性差异（χ2分别为0.21和0.618,P＞0.05）,Youden指数分别为0.91和0.92;对并殖吸虫感染者全血检测的交叉反应率为43.8%（7/16）,其他寄生虫感染者全血检测的交叉反应率为0（0/29）.研究结果显示WB-DLIA与ELISA具有相似的敏感性和特异性,但前者无需分离血清和任何仪器,结果观察清晰,操作更加简便和快速,且易于保存运输,更加适于临床检测、基层推广和大规模现场筛查.

A rapid whole-blood dipstick latex immunochromatography assay （WB-DLIA） was established based on the principle of indirect immunoassay for detecting the antibody in blood sample from patients with schistosomiasis. The dipstick used soluble egg antigen of Schistosoma japonicum and mouse anti-human monoclonal antibody labeled with latex as color probe, and the outcomes of WB-DLIA were compared with those of ELISA to assess the practical value of the established method. The positive coincidences of WB-DLIA and ELISA in detecting 54 whole-blood samples from stool-positive patients were 94. 4% （51/54） and 96. 3% （52/54）, respectively, with no significant difference （χ2 = 0. 21, P 〉 0. 05 ）. The negative coincidences in detecting 261 whole-blood sample from healthy people were 96.6% （252/261） and 95.4% （249/ 261 ） , respectively, also with no significant difference （χ2 = 0. 618, P 〉 0. 05 ）. The Youden＇s indexes were 0. 91 and 0. 92, respectively. The cross-reaction rate in 16 samples from patients with paragonimiasis was 43.8% （7/16） both in WB-DLIA and ELISA, and no cross-reaction occurred in 29 whole-blood samples from patients with other parasitic diseases. The results of present study indicated that WB-DLIA possesses the same sensitivity and specificity as ELISA, with the advantages of both practical and simple, for no any other equipment and serum separation needed. WB-DLIA is suitable for spread application at grass-roots level and for general field screening of schistosomiasis.