Goh K H, Ng M L, Thean E T, Khalid B A
Department of Biochemistry, Faculty of Medicine, Universiti Kebangsaan Malaysia, Kuala Lumpur.
Med J Malaysia. 1992 Dec;47(4):248-60.
A supersensitive ELISA was developed for measurement of thyroid-stimulating hormone (TSH) concentrations in serum using in-house rabbit polyclonal antisera and a commercial monoclonal antibody. The assay was optimised and validated by recovery, linearity and cross-reactivity experiments and further compared to other available assays and EQAS samples. Good precision was obtained with a working assay range of 0.2 to 100 mIU/L with < 10% coefficient of variation (CV) for both intra and interassay. The assay is highly sensitive and specific with a minimum detectable limit of 0.07 mIU/L and negligible cross-reactivities against LH, FSH, HCG and other pituitary peptides. Good correlations were obtained when compared to Abbott hTSH EIA (r = 0.993; p < 0.001; n = 85) and NETRIA IRMA (r + 0.995; p < 0.001; n = 76). The normal reference range established was 0.4 to 4.0 mIU/L (n = 76). TSH levels in serum of thyrotoxic patients (n = 83) were significantly lower (0.07 to 0.20 mIU/L, p < 0.0001) and completely distinct from normal values thereby obviating the requirement of a TRH-stimulation test. Stability studies showed that coated wells can be stored at 4 degrees C for at least 2 months. This highly sensitive in-house hTSH ELISA which is cheap, stable and readily available is useful for diagnosis and management of patients with various thyroid disorders.
利用自制兔多克隆抗血清和一种商用单克隆抗体，开发了一种超灵敏酶联免疫吸附测定法（ELISA），用于测定血清中促甲状腺激素（TSH）的浓度。通过回收率、线性和交叉反应实验对该测定法进行了优化和验证，并进一步与其他现有测定法和室间质量评价计划（EQAS）样本进行了比较。在0.2至100 mIU/L的工作测定范围内获得了良好的精密度，批内和批间变异系数（CV）均<10%。该测定法具有高灵敏度和特异性，最低检测限为0.07 mIU/L，与促黄体生成素（LH）、促卵泡生成素（FSH）、人绒毛膜促性腺激素（HCG）和其他垂体肽的交叉反应可忽略不计。与雅培hTSH酶免疫测定法（r = 0.993；p < 0.001；n = 85）和NETRIA免疫放射测定法（r = 0.995；p < 0.001；n = 76）相比，获得了良好的相关性。建立的正常参考范围为0.4至4.0 mIU/L（n = 76）。甲状腺毒症患者（n = 83）血清中的TSH水平显著降低（0.07至0.20 mIU/L，p < 0.0001），与正常值完全不同，因此无需进行促甲状腺激素释放激素（TRH）刺激试验。稳定性研究表明，包被孔可在4℃下储存至少2个月。这种高度灵敏的自制hTSH ELISA价格便宜、稳定且易于获得，可用于各种甲状腺疾病患者的诊断和管理。
