Requejo H I, Alkmin M das G, Almeida R G, Casagrande S T, Cocozza A M, Lotufo J P, Waetge A R, Rodrigues J C
Seção de Imunologia, Instituto Adolfo Lutz, São Paulo, Brasil.
Rev Inst Med Trop Sao Paulo. 1994 Nov-Dec;36(6):531-7. doi: 10.1590/s0036-46651994000600010.
A dot-enzyme-linked immunosorbent assay (Dot-ELISA) for pneumococcal antigen detection was standardized in view of the need for a rapid and accurate immunodiagnosis of acute pneumococcal pneumonia. A total of 442 pleural fluid effusion samples (PFES) from children with clinical and laboratory diagnoses of acute bacterial pneumonia, plus 38 control PFES from tuberculosis patients and 20 negative control serum samples from healthy children were evaluated by Dot-ELISA. The samples were previously treated with 0.1M EDTA pH 7.5 at 90 degrees C for 10 min and dotted on nitrocellulose membrane. Pneumococcal omniserum diluted at 1:200 was employed in this assay for antigen detection. When compared with standard bacterial culture, counterimmunoelectrophoresis and latex agglutination techniques, the Dot-ELISA results showed relative indices of 0.940 to sensitivity, 0.830 to specificity and 0.760 to agreement. Pneumococcal omniserum proved to be an optimal polyvalent antiserum for the detection of pneumococcal antigen by Dot-ELISA. Dot-ELISA proved to be a practical alternative technique for the diagnosis of pneumococcal pneumonia.
鉴于对急性肺炎球菌肺炎进行快速准确免疫诊断的需求，对一种用于检测肺炎球菌抗原的斑点酶联免疫吸附测定法（Dot-ELISA）进行了标准化。通过Dot-ELISA对442例临床和实验室诊断为急性细菌性肺炎患儿的胸腔积液样本（PFES）、38例结核病患者的对照PFES以及20例健康儿童的阴性对照血清样本进行了评估。样本预先在90℃下用0.1M pH 7.5的乙二胺四乙酸（EDTA）处理10分钟，然后点样于硝酸纤维素膜上。本试验采用稀释至1:200的肺炎球菌全价血清进行抗原检测。与标准细菌培养、对流免疫电泳和乳胶凝集技术相比，Dot-ELISA结果的敏感性相对指数为0.940，特异性相对指数为0.830，一致性相对指数为0.760。肺炎球菌全价血清被证明是通过Dot-ELISA检测肺炎球菌抗原的最佳多价抗血清。Dot-ELISA被证明是诊断肺炎球菌肺炎的一种实用替代技术。
