Detection of antibody IgG to HIV‐1 in urine by sensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) using recombinant proteins as antigens for diagnosis of HIV‐1 infection
Detection of antibody IgG to HIV‐1 in urine by sensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) using recombinant proteins as antigens for diagnosis of HIV‐1 infection
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使用重组蛋白作为抗原，通过敏感酶免疫分析（免疫复合物转移酶免疫分析）检测尿液中 HIV-1 抗体 IgG，诊断 HIV-1 感染
DOI：
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发表时间：
1994
期刊：
影响因子：
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通讯作者：
E. Ishikawa
中科院分区：
文献类型：
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作者：
S. Hashida;K. Hirota;K. Hashinaka;A. Saitoh;A. Nakata;H. Shinagawa;S. Oka;K. Shimada;J. Mimaya;S. Matsushita;E. Ishikawa
为了诊断HIV-1感染，尝试使用重组逆转录酶（RT）和p17作为抗原，通过灵敏的酶免疫测定（免疫复合物转移酶免疫测定）检测尿液中的抗HIV-1 IgG。尿液中的抗HIV-1 IgG同时与2，4-二硝基苯-牛血清白蛋白-重组蛋白偶联物和重组蛋白酶偶联物反应。用作标记的酶是用于RT的辣根过氧化物酶和用于p17的大肠杆菌β-D-半乳糖苷酶。将形成的由三种组分组成的复合物捕获到包被有亲和纯化的（抗2，4-二硝基苯基）IgG的聚苯乙烯球上，用∈N-2，4-二硝基苯基-L-赖氨酸洗脱，并转移到包被有亲和纯化的（抗人IgG γ链）IgG的聚苯乙烯球上。最后，通过荧光测定法测定结合的酶活性。从100名血清阴性受试者和70名血清阳性受试者中采集尿样。对未浓缩尿样的敏感性和特异性均为100%。通过用过量抗原预孵育尿样确认阳性。两种抗原之一的阳性和阴性可以用另一种抗原证实。低信号阳性可通过尿样浓度确认。通过使用不同抗原的免疫复合物转移酶免疫测定法检测尿液中的抗HIV-1 IgG将使HIV-1感染的诊断成为可能。© 1993 Wiley利斯公司
For diagnosis of HIV‐1 infection, attempts were made to detect anti‐HIV‐1 IgG in urine by sensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) using recombinant reverse transcriptase (RT) and p 17 as antigens. Anti‐HIV‐1 IgG in urine was reacted simultaneously with 2,4‐dinitrophenyl‐bovine serum albumin‐recombinant protein conjugate and recombinant proteinenzyme conjugate. The enzymes used as labels were horseradish peroxidase for RT and Escherichia coli β‐D‐galactosidase for p17. 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 ∈N‐2,4‐dinitrophenyl‐L‐lysine and transferred to polystyrene balls coated with affinity‐purified (anti‐human IgG γ‐chain) IgG. Finally, bound enzyme activity was assayed by fluorometry. Urine samples were collected from 100 seronegative subjects and 70 seropositive subjects. The sensitivity and specificity were both 100% with unconcentrated urine samples. The positivity was confirmed by preincubation of urine samples with excess of the antigens. The positivity and negativity with one of the two antigens could be confirmed with the other antigen. The positivity with low signals could be confirmed by concentration of urine samples. Detection of anti‐HIV‐1 IgG in urine by the immune complex transfer enzyme immunoassay using different antigens would make diagnosis of HIV‐1 infection possible. © 1993 Wiley‐Liss, Inc.