增强化学发光酶免疫法测定血清促甲状腺激素方法的建立及初步应用

Establishment and application of a new chemiluminescence enzyme immunoassay to determine human thyrotropin in serum

目的进一步提高血清促甲状腺激素（ＴＳＨ）检测的敏感性。方法通过正交试验，并经方差分析，获得增强发光酶免疫检测法的最佳实验条件，将其与ＴＳＨ酶联免疫吸附测定（ＥＬＩＳＡ）试剂盒相结合，建立ＴＳＨ的增强化学发光酶免疫检测法，并用此方法对５９例健康儿童和４９例甲状腺疾病患儿进行了血清ＴＳＨ的检测。结果该方法检测人血清ＴＳＨ的灵敏度为０．０１ｍＩＵ／Ｌ。５９例健康儿童ＴＳＨ的９０％正常值范围为０．７４～８．５３ｍＩＵ／Ｌ；１７例甲状腺功能亢进患儿ＴＳＨ值范围为０．０２～０．４０ｍＩＵ／Ｌ，无一例与正常值重叠；１６例甲状腺功能正常的弥漫性甲状腺肿患儿ＴＳＨ值均在正常范围上限；１６例甲状腺功能低下患儿的ＴＳＨ值均大于３０ｍＩＵ／Ｌ。结论该方法与ＲＩＡ、ＥＬＩＳＡ比较，有良好的相关性，可基本分清甲状腺功能亢进、低下、正常三种状态，适于临床甲状腺功能的评估。

Objective To improve the sensitivity of measurement of TSH in serum. Methods The authors developed a new enhanced chemiluminescence enzyme immunoassay (ECLIA), which combined luminol H 2O 2 HRP system enhanced by para iodophenol(PIP) with TSH ELISA test kit. The optimal experimental conditions of the enhanced chemiluminescence were obtained by orthogonal design and analysis of variance. Results The sensitivity of the assay was 0.01mIU/L. The intra and inter coefficients of variations were 3.95% (range 1.37%～7.43%) and 6.98% (range 3.99%～10.57%), respectively. The recovery rate was 97.57%(range 93.11%～102.48%), and there was no significant cross reaction with LH, FSH or HCG. Results of the ECLIA correlated well with those of RIA or ELISA ( r =0.94, n =35, P <0.001 and r =0.98, n =22, P < 0.001). The 0.90 confidence interval of TSH for normal children by ECLIA was 0.74～8.53 mIU/L ( n =59). Circulating TSH concentrations in 17 hyperthyroids were 0.02～0.40 mIU/L, with no one in the normal range; in 16 euthyroids were all in the normal range, but concentrated on the upper limit of normal; in 16 hypothyroids the values were beyond 30 mIU/L. Conclusion The ECLIA method can discriminate hyperthyroidism, hypothyroidism and euthyroidism.