da Silva M V, Nakamura P M, Camargo E D, Batista L, Vaz A J, Brandão A P, Ferreira A W
Instituto Adolfo Lutz, São Paulo, Brasil.
Rev Inst Med Trop Sao Paulo. 1994 Sep-Oct;36(5):475-8. doi: 10.1590/s0036-46651994000500014.
In order to improve the diagnosis of human leptospirosis, we standardized the dot-ELISA for the search of specific IgM antibodies in saliva. Saliva and serum samples were collected simultaneously from 20 patients with the icterohemorrhagic form of the disease, from 10 patients with other pathologies and from 5 negative controls. Leptospires of serovars icterohaemorrhagiae, canicola, hebdomadis, brasiliensis and cynopteri grown in EMJH medium and mixed together in equal volumes, were used as antigen at individual protein concentration of 0.2 micrograms/microliters. In the solid phase of the test we used polyester fabric impregnated with N-methylolacrylamide resin. The antigen volume for each test was 1 microliter, the saliva volume was 8 microliters, and the volume of peroxidase-labelled anti-human IgM conjugate was 30 microliters. A visual reading was taken after development in freshly prepared chromogen solution. In contrast to the classic nitrocellulose membrane support, the fabric support is easy to obtain and to handle. Saliva can be collected directly onto the support, a fact that facilitates the method and reduces the expenses and risks related to blood processing.
为提高人钩端螺旋体病的诊断水平，我们对用于检测唾液中特异性IgM抗体的斑点酶联免疫吸附试验（dot-ELISA）进行了标准化。同时收集了20例黄疸出血型钩端螺旋体病患者、10例患有其他病症的患者以及5例阴性对照者的唾液和血清样本。将在EMJH培养基中培养并等体积混合的黄疸出血型、犬型、七日热型、巴西型和犬蝠型钩端螺旋体用作抗原，其各自的蛋白质浓度为0.2微克/微升。在试验的固相阶段，我们使用了浸渍有N-羟甲基丙烯酰胺树脂的聚酯织物。每次试验的抗原体积为1微升，唾液体积为8微升，过氧化物酶标记的抗人IgM结合物体积为30微升。在新鲜配制的显色溶液中显色后进行目视读数。与传统的硝酸纤维素膜载体相比，织物载体易于获取和操作。唾液可直接收集到载体上，这一事实简化了方法，并降低了与血液处理相关的费用和风险。
