Immunoreactive insulin in diabetes mellitus patient sera detected by ultrasensitive ELISA with thio-NAD cycling
Immunoreactive insulin in diabetes mellitus patient sera detected by ultrasensitive ELISA with thio-NAD cycling
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DOI：
10.2144/000114355
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发表时间：
2015-12-01
期刊：
影响因子：
2.7
通讯作者：
Miura, Toshiaki
中科院分区：
文献类型：
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作者：
Ito, Etsuro;Kaneda, Mugiho;Miura, Toshiaki
为了最大限度地减少患者的痛苦，应收集尽可能少的血液用于诊断和疾病监测。在评估糖尿病（DM）患者的胰岛素分泌能力和胰岛素抵抗时，增加胰岛素测定免疫敏感性将减少检测所需的血样体积。在这里，我们提出了一种超灵敏的ELISA加上硫代NAD循环来测量血清中的免疫反应性胰岛素。检测仅需5 mL血清，检测限（LOD）为10-16摩尔/次。另外的回收试验证实该方法可以检测血清中的胰岛素。市售免疫反应性胰岛素试剂盒与我们使用相同市售参比品的超灵敏ELISA之间的比较证明了良好的数据相关性，为测定准确度提供了进一步证据。总之，这些结果表明，我们的超灵敏ELISA可能是一个强大的工具，不仅在诊断和治疗糖尿病，而且在未来的许多其他疾病。
To minimize patient suffering, the smallest possible volume of blood should be collected for diagnosis and disease monitoring. When estimating insulin secretion capacity and resistance to insulin in diabetes mellitus (DM), increasing insulin assay immunosensitivity would reduce the blood sample volume required for testing. Here we present an ultrasensitive ELISA coupled with thio-NAD cycling to measure immunoreactive insulin in blood serum. Only 5 mL of serum was required for testing, with a limit of detection (LOD) for the assay of 10-16 moles/assay. Additional recovery tests confirmed this method can detect insulin in sera. Comparisons between a commercially available immunoreactive insulin kit and our ultrasensitive ELISA using the same commercially available reference demonstrated good data correlation, providing further evidence of assay accuracy. Together, these results demonstrate our ultrasensitive ELISA could be a powerful tool in the diagnosis and treatment of not only DM but also many other diseases in the future.