Hashida S, Hirota K, Kohno T, Ishikawa E
Department of Biochemistry, Medical College of Miyazaki, Japan.
J Clin Lab Anal. 1994;8(3):149-56. doi: 10.1002/jcla.1860080307.
Antibody IgG to human T-cell leukemia virus type I (HTLV-I) in urine was detected by a sensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) using a synthetic peptide, Cys-env gp46(188-224), as antigen, the sensitivity and specificity of which were 100 and 98.5%, respectively, using serum samples. Anti-HTLV-I IgG in urine was reacted simultaneously with 2,4-dinitrophenyl-bovine serum albumin-Cys-env gp46(188-224) conjugate and Cys-env gp46(188-224)-beta-D-galactosidase (Escherichia coli) conjugate. The complex formed, consisting of the three components, was trapped onto polystyrene balls coated with affinity-purified (anti-2,4-dinitrophenyl group) IgG, eluted with epsilon N-2,4-dinitrophenyl-L-lysine and transferred to polystyrene balls coated with affinity-purified (anti-human IgG gamma-chain) IgG. Finally, bound beta-D-galactosidase activity was assayed by fluorometry. Thirty-one urine samples from seropositive subjects and 100 urine samples from seronegative subjects were tested. The sensitivity and specificity were 87 and 100%, respectively, with unconcentrated urine samples and 94 and 100%, respectively, with approximately 10-fold concentrated urine samples. These results were superior to those by the conventional ELISA and gelatin particle agglutination test.
采用合成肽Cys-env gp46(188 - 224)作为抗原,通过灵敏的酶免疫测定法(免疫复合物转移酶免疫测定法)检测尿液中针对人I型T细胞白血病病毒(HTLV - I)的抗体IgG。使用血清样本时,该方法的灵敏度和特异性分别为100%和98.5%。尿液中的抗HTLV - I IgG同时与2,4 - 二硝基苯基 - 牛血清白蛋白 - Cys-env gp46(188 - 224)偶联物和Cys-env gp46(188 - 224)-β-D-半乳糖苷酶(大肠杆菌)偶联物反应。由这三种成分形成的复合物被捕获到包被有亲和纯化的(抗2,4 - 二硝基苯基基团)IgG的聚苯乙烯球上,用ε-N-2,4 - 二硝基苯基-L-赖氨酸洗脱,然后转移到包被有亲和纯化的(抗人IgGγ链)IgG的聚苯乙烯球上。最后,通过荧光测定法检测结合的β-D-半乳糖苷酶活性。检测了31份血清阳性受试者的尿液样本和100份血清阴性受试者的尿液样本。未浓缩尿液样本的灵敏度和特异性分别为87%和100%,约10倍浓缩尿液样本的灵敏度和特异性分别为94%和100%。这些结果优于传统酶联免疫吸附测定法和明胶颗粒凝集试验的结果。