Kohno T, Sakoda I, Ishikawa E
Department of Biochemistry, Medical College of Miyazaki, Japan.
J Clin Lab Anal. 1992;6(3):162-9. doi: 10.1002/jcla.1860060311.
A sensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) for (antihuman T-cell leukemia virus type I) IgG (anti-HTLV-I IgG) in serum using a synthetic peptide, Ala-Cys-env gp46(237-262), of HTLV-I is described. Anti-HTLV-I IgG in test serum, which had been incubated with excess of inactive beta-D-galactosidase to eliminate interference by anti-beta-D-galactosidase antibodies, was reacted simultaneously with 2,4-dinitrophenyl-bovine serum albumin-Ala-Cys-env gp46(237-262) conjugate and Ala-Cys-env gp46(237-262)-beta-D-galactosidase conjugate. The complex formed consisting of the three components was trapped onto polystyrene balls coated with affinity-purified (anti-2,4-dinitrophenyl group) IgG. After washing to eliminate nonspecific IgG in the test serum and excess of the beta-D-galactosidase conjugate, the complex was eluted from the polystyrene balls with epsilon N-2,4-dinitrophenyl-L-lysine and transferred to polystyrene balls coated with affinity-purified (anti-human IgG gamma-chain) IgG. beta-D-galactosidase activity bound to the (anti-human IgG gamma-chain) IgG-coated polystyrene balls was assayed by fluorometry. This assay was more sensitive than other methods using HTLV-I as antigen, and most negative and positive sera were discriminated. However, some results appeared to be false positive or false negative, and the peptide, Ala-Cys-env gp46(237-262), was suggested to be useful, in combination with other peptides, for improving the reliability of serodiagnosis by separately demonstrating antibodies against as many different epitopes of HTLV-I as possible.
描述了一种使用人嗜T淋巴细胞病毒I型(HTLV-I)的合成肽Ala-Cys-env gp46(237 - 262)检测血清中抗人嗜T淋巴细胞病毒I型IgG(抗HTLV-I IgG)的灵敏酶免疫测定法(免疫复合物转移酶免疫测定法)。将检测血清中的抗HTLV-I IgG与过量的无活性β-D-半乳糖苷酶孵育以消除抗β-D-半乳糖苷酶抗体的干扰后,使其与2,4-二硝基苯基-牛血清白蛋白-Ala-Cys-env gp46(237 - 262)偶联物和Ala-Cys-env gp46(237 - 262)-β-D-半乳糖苷酶偶联物同时反应。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。洗涤以去除检测血清中的非特异性IgG和过量的β-D-半乳糖苷酶偶联物后,用ε-N-2,4-二硝基苯基-L-赖氨酸从聚苯乙烯球上洗脱复合物,并转移到包被有亲和纯化的(抗人IgGγ链)IgG的聚苯乙烯球上。通过荧光测定法检测与(抗人IgGγ链)IgG包被的聚苯乙烯球结合的β-D-半乳糖苷酶活性。该测定法比使用HTLV-I作为抗原的其他方法更灵敏,并且大多数阴性和阳性血清都能被区分。然而,一些结果似乎为假阳性或假阴性,并且有人提出肽Ala-Cys-env gp46(237 - 262)与其他肽联合使用,通过尽可能分别显示针对HTLV-I许多不同表位的抗体,对于提高血清学诊断的可靠性是有用的。