Bergman C, Crisci C D, Jinnouchi H, Oehling A
Allergol Immunopathol (Madr). 1979 Jul-Aug;7(4):283-8.
A new application of the Sepharose bead immunofluorescence test for detection of allergen-specific IgE and IgG antibodies is described. Allergen extracts of four different grass pollens were coupled to CNBr-activated Sepharose 4B. Twenty normal and allergic sera were incubated with the allergen-coupled beads, washed and incubated with fluorescence-conjugated anti-gamma E and G globulins. After washing and staining with 0.5% trypan blue, the percentage of fluorescent beads was detected by fluorescence microscopy. In the IgG/anti IgG system, the smallest amount of IgG demonstrated was 20ng/ml; in the IgE/anti-IgE system it was 40ng/ml. Independent examination of tests gave a mean difference between observations of 4.1%. Reproducibility was also very good (var. coeff = 18%). The number of beads stained with IgE correlated well with intensity of skin reactions to the same extracts (r = 0.64; p = 5 x 10-7), the percentage of IgG stained beads being independent of skin reactivity. The Sepharose-IgE test allowed clear distinction between allergic and normal sera (p = 4 x 10-7), while the Sepharose IgG test did not distinguish between them at the time of diagnosis. However, the number of beads stained with IgG significantly increased in patients undergoing immunotherapy (p = 3.8 x 10-3). The Sepharose bead immunofluorescence test requires a very small amount of materials, is highly sensitive and easy to handle. It may be valuable in the "in vitro" diagnosis of grass pollen allergy and useful in evaluating immunotherapy.
本文描述了琼脂糖珠免疫荧光试验在检测变应原特异性IgE和IgG抗体方面的一种新应用。将四种不同草花粉的变应原提取物偶联到溴化氰活化的琼脂糖4B上。将20份正常血清和过敏血清与变应原偶联的珠子孵育，洗涤后再与荧光标记的抗γE和G球蛋白孵育。洗涤并用0.5%台盼蓝染色后，通过荧光显微镜检测荧光珠的百分比。在IgG/抗IgG系统中，检测到的最小IgG量为20ng/ml；在IgE/抗IgE系统中为40ng/ml。对试验进行独立检测得出观察结果之间的平均差异为4.1%。重复性也非常好（变异系数=18%）。用IgE染色的珠子数量与对相同提取物的皮肤反应强度密切相关（r = 0.64；p = 5×10-7），而用IgG染色的珠子百分比与皮肤反应性无关。琼脂糖-IgE试验能够清晰地区分过敏血清和正常血清（p = 4×10-7），而在诊断时琼脂糖IgG试验无法区分它们。然而，接受免疫治疗的患者中用IgG染色的珠子数量显著增加（p = 3.8×10-3）。琼脂糖珠免疫荧光试验所需材料非常少，灵敏度高且易于操作。它在草花粉过敏的“体外”诊断中可能具有重要价值，并且有助于评估免疫治疗效果。
