Dahlberg T, Branefors P
J Clin Microbiol. 1980 Aug;12(2):185-92. doi: 10.1128/jcm.12.2.185-192.1980.
The enzyme-linked immunosorbent assay (ELISA) was elaborated for the detection of immunoglobulin M (IgM) and IgG antibodies against capsular and O antigens of Haemophilus influenzae. Purified capsular polysaccharide and lipopolysaccharide were used as antigens, with optimal coating concentrations being about 50 and 100 micrograms/ml, respectively. The antibody content was expressed as the highest serum dilution (-log10) showing an absorbance of 0.2 above the background level. The titers of hyperimmune sera (reference sera) ranged between 5 and 7 -log10. The sensitivity of the method was about 80 ng/ml with regard to anticapsular antibodies and 3 to 5 ng/ml with regard to anti-lipopolysaccharide antibodies. For detection of antibodies against capsular polysaccharide in sera obtained after primary immunization, ELISA was about 100-fold more sensitive than the indirect hemagglutination assay, whereas in hyperimmune sera, ELISA was about 10-fold more sensitive than the indirect hemagglutination assay. The sensitivity of ELISA for detecting anticapsular antibodies after primary and booster immunizations was 50-fold higher than that of the bactericidal assay using capsulated bacteria, whereas the sensitivity of the two methods was the same when hyperimmune sera were tested. ELISA performed with lipopolysaccharide as the antigen was about 50- and 150-fold more sensitive than the complement fixation and bactericidal assays tested with noncapsulated variants after primary injection and hyperimmunization, respectively.
酶联免疫吸附测定法（ELISA）被用于检测针对流感嗜血杆菌荚膜和O抗原的免疫球蛋白M（IgM）和IgG抗体。纯化的荚膜多糖和脂多糖被用作抗原，最佳包被浓度分别约为50微克/毫升和100微克/毫升。抗体含量以高于背景水平吸光度为0.2时的最高血清稀释度（-log10）表示。超免疫血清（参考血清）的滴度范围在5至7 -log10之间。该方法对抗荚膜抗体的灵敏度约为80纳克/毫升，对抗脂多糖抗体的灵敏度为3至5纳克/毫升。对于检测初次免疫后获得的血清中针对荚膜多糖的抗体，ELISA比间接血凝试验灵敏约100倍，而在超免疫血清中，ELISA比间接血凝试验灵敏约10倍。ELISA检测初次免疫和加强免疫后抗荚膜抗体的灵敏度比使用有荚膜细菌的杀菌试验高50倍，而检测超免疫血清时两种方法的灵敏度相同。以脂多糖为抗原进行的ELISA在初次注射和超免疫后分别比用无荚膜变体检测的补体结合试验和杀菌试验灵敏约50倍和150倍。
