Use of microplates and fluororeader for ultrasensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) of anti‐HTLV‐I IgG
Use of microplates and fluororeader for ultrasensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) of anti‐HTLV‐I IgG
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使用微孔板和荧光读数器进行抗 HTLV-I IgG 的超灵敏酶免疫测定（免疫复合物转移酶免疫测定）
DOI：
10.1002/jcla.1860080512
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发表时间：
1994
影响因子：
2.7
通讯作者：
E. Ishikawa
中科院分区：
文献类型：
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作者：
S. Ishikawa;S. Hashida;T. Kohno;E. Ishikawa
以前，描述了一种超灵敏的固相酶免疫测定法（免疫复合物转移酶免疫测定法），用于检测血清中低于常规方法可检测水平的低水平抗HTLV-I IgG。该方法以聚苯乙烯球为固相，用镊子将固相从试管转移到试管。这不仅冗长乏味，而且还会导致残留的假阳性，除非每次转移聚苯乙烯球后仔细清洗镊子尖端。使用荧光分光光度计通过荧光测定法逐一测量许多样品的结合酶活性。因此，许多样品的测定是困难的。在本研究中，使用微孔板和荧光阅读器代替试管和荧光分光光度计。通过将微板倒置在含有聚苯乙烯球的微板上，并将两个板一起倒置，将聚苯乙烯球快速且容易地从一个孔转移到另一个孔。不使用镊子转移聚苯乙烯球，以最大限度地降低假阳性的可能性。通过使用荧光读数器在1分钟内测量96个样品的结合酶活性的荧光强度。因此，尽管灵敏度在一定程度上降低，但测试许多样品变得容易。© 1994 Wiley利斯公司
Previously, an ultrasensitive solid phase enzyme immunoassay (immune complex transfer enzyme immunoassay) was described to detect low levels of anti‐HTLV‐I IgG in serum below those detectable by conventional methods. In this method, polystyrene balls as solid phase were transferred from test tube to test tube with tweezers. This was not only tedious but also causative of false‐positivity by carryover, unless tips of the tweezers were washed carefully after each transfer of polystyrene balls. Bound enzyme activities for many samples were measured one by one by fluorometry using a spectrofluorophotometer. As a result, the assay of many samples was difficult. In the present study, microplates and a fluororeaderwere used in place of test tubes and a spectrofluorophotometer. Polystyrene balls were transferred quickly and easily from well to well by placing a microplate upside down on that containing polystyrene balls, and turning the two plates together upside down. Tweezers were not used for transfer of polystyrene balls, minimizing the possibility of false‐positivity. Fluorescence intensities of bound enzyme activities for 96 samples were measured within a minute by using a fluororeader. Thus, it became easy to test many samples, although the sensitivity was lowered to some extent. © 1994 Wiley‐Liss, Inc.
Hashida,S.：“使用重组蛋白作为诊断 HIV-1 感染的抗原，通过敏感酶免疫测定（免疫复合物转移酶免疫测定）检测尿液中 HIV-1 抗体 IgG”J.Clin.Lab.Anal.7。
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Takeyuki Kohno 等人：“使用来自埃希氏菌的 β-D-半乳糖苷酶来消除血清中抗甲状腺球蛋白 IgG 的免疫复合物转移酶免疫测定中抗 β-D-半乳糖苷酶抗体的干扰大肠杆菌
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