Slater J E, Trybul D E
Department of Allergy and Immunology, Children's National Medical Center, Washington, D.C. 20010.
J Allergy Clin Immunol. 1994 May;93(5):825-30. doi: 10.1016/0091-6749(94)90372-7.
Protein antigens in latex products can cause type I reactions. In the past antigens have been measured by protein assays, high-performance liquid chromatography, RAST inhibition, and skin tests. We examined the use of a mouse monoclonal antibody (CRI-C) to latex in the detection of latex antigens.
CRI-C was raised by standard techniques after immunization of a BALB/c mouse with ammoniated latex. Medical gloves were extracted and assayed with: (1) standard protein assays, (2) RAST inhibition assays with sera from health care workers allergic to latex and patients with spina bifida, (3) an ELISA with a biotinylated form of CRI-C (BiC). Reference proteins included bovine serum albumin for the protein assays and nonammoniated latex and affinity-purified C antigen for the immunoassays.
Among the protein assays, the best correlation was between the Bradford and bicinchoninic acid assays. In absolute numbers the Bradford assay produced the lowest results, and OD280 the highest. The OD280, BCA, and Bradford methods "detected" protein in vinyl gloves. The results of RAST inhibition and BiC ELISA correlated with the protein assays. These immunoassays appeared to be more specific than the protein assays.
The BiC ELISA is an easy and reproducible in vitro assay of relevant latex antigen. Clinical correlation will be required for validation.
乳胶制品中的蛋白质抗原可引发I型反应。过去,抗原检测采用蛋白质测定法、高效液相色谱法、放射性变应原吸附抑制试验(RAST抑制试验)和皮肤试验。我们研究了使用小鼠抗乳胶单克隆抗体(CRI-C)检测乳胶抗原的情况。
用氨化乳胶免疫BALB/c小鼠后,通过标准技术制备CRI-C。提取医用手套并进行以下检测:(1)标准蛋白质测定法;(2)对乳胶过敏的医护人员和脊柱裂患者血清的RAST抑制试验;(3)生物素化形式的CRI-C(BiC)酶联免疫吸附测定(ELISA)。参考蛋白质包括用于蛋白质测定的牛血清白蛋白以及用于免疫测定的未氨化乳胶和亲和纯化的C抗原。
在蛋白质测定法中,考马斯亮蓝法和二喹啉甲酸法之间的相关性最佳。就绝对数值而言,考马斯亮蓝法结果最低,280nm吸光度法结果最高。280nm吸光度法、二喹啉甲酸法和考马斯亮蓝法在乙烯基手套中“检测”到了蛋白质。RAST抑制试验和BiC ELISA的结果与蛋白质测定法相关。这些免疫测定法似乎比蛋白质测定法更具特异性。
BiC ELISA是一种简便且可重复的体外检测相关乳胶抗原的方法。需进行临床相关性研究以验证其有效性。