组分抗原斑点金标免疫渗滤法检测血吸虫短程抗体的研究

目的建立一种可用于检测血吸虫病人短程抗体的斑点金标免疫渗滤法（dot immuno-gold filtration assay,DIG-FA）。方法将血吸虫可溶性虫卵组分抗原（107～121 kDa）包被于混纤膜上,以胶体金标记的抗人IgG为二抗,建立组分抗原-DIGFA。用该法检测慢性血吸虫病人、健康人、卫氏并殖吸虫病人和华支睾吸虫病人血清。同时与血吸虫可溶性虫卵抗原（SEA）-DIGFA比较。结果检测54份慢性血吸虫病人血清,组分抗原-DIGFA敏感性为94.4%,SEA-DIGFA敏感性为96.3%,两种方法检测50份正常人血清的特异性均为100%。检测19份卫氏并殖吸虫病人血清和30份华支睾吸虫病人血清,SEA-DIGFA敏感性分别为26.3%和10.0%,组分抗原-DIGFA均未检出阳性反应。结论应用组分抗原-DIGFA检测血吸虫短程抗体具有较高的敏感性和特异性,且与其他吸虫病几乎无交叉反应,特异性优于SEA-DIGFA。

Study on detection of short term antibodies of schistosomiasis with dot immuno-gold filtration assay(DIGFA)

This study was to establish and evaluate the dot immuno-gold filtration assay（DIGFA） for detecting short term anti- bodies in patients＇ serum with fraction antigen of SEA of Schistosomajaponicum. SEA and the fraction antigen （ 107 - 121 kDa） of SEA were prepared. One μl of fraction antigen of SEA was dotted on the mixed fiber membrane, and 20 μl of detected serum sample was placed on the dotted paper. After washing with 0.01 mol/L PBS-T, 100 μl of diluted anti-human IgG conjugated with colloidal gold was placed on the paper as the second antibody. If the dot was red, the sample was recorded as positive. The results showed that 51of 54 patients＂ sera of schistosomiasis were positive by fraction-DIGFA, the positive rate was 94.4% ; 52 of 54 sera were positive by SEA-DIGFA, the positive rate was 96.3%. Fifty of healthy people＇s sera were negative by both methods. Nineteen paragonimiasis sera and 30 clonorchiasis sera were negative by fraction-DIGFA, but 26.3% （5/19） and 10.0% （3/30） of them were oositive by SEA-DIGFA resoectivelv. It indicated that fraction-DIGFA had high sensitivity and high specificity.