硫酸脫氫表雄酮化學(xué)發(fā)光微粒子免疫法定量測定試劑的研制

黃明聰　張曉琍　林光華　翁祖星　孫旭東　葛勝祥　張　軍★

硫酸脫氫表雄酮化學(xué)發(fā)光微粒子免疫法定量測定試劑的研制

黃明聰1張曉琍2林光華2翁祖星1孫旭東1葛勝祥3張軍3★

［摘要］目的研制硫酸脫氫表雄酮化學(xué)發(fā)光免疫定量檢測試劑。方法利用硫酸脫氫表雄酮人工完全抗原免疫小鼠，通過雜交瘤技術(shù)制備特異性抗硫酸脫氫表雄酮單克隆抗體，采用競爭抑制法建立硫酸脫氫表雄酮化學(xué)發(fā)光免疫定量檢測試劑。結(jié)果篩選獲得了27株穩(wěn)定分泌抗硫酸脫氫表雄酮的單克隆抗體細(xì)胞株，建立了化學(xué)發(fā)光微粒子免疫法定量測定硫酸脫氫表雄酮的試劑盒雛形，與雅培公司的硫酸脫氫表雄酮定量檢測試劑在檢測臨床標(biāo)本上的相關(guān)系數(shù)r達(dá)0．99以上。結(jié)論本研究為國產(chǎn)化硫酸脫氫表雄酮化學(xué)發(fā)光微粒子免疫法定量測定試劑盒的研發(fā)奠定了基礎(chǔ)。

［關(guān)鍵詞］硫酸脫氫表雄酮；化學(xué)發(fā)光微粒子免疫法；定量試劑盒

作者單位：1．廈門萬泰凱瑞生物技術(shù)有限公司，福建，廈門361022 2．福建醫(yī)科大學(xué)附屬協(xié)和醫(yī)院檢驗(yàn)科，福建，福州350001 3．廈門大學(xué)國家傳染病診斷試劑與疫苗工程技術(shù)研究中心，福建，廈門361102

注：黃明聰和張曉琍為并列第一作者

Development of chemiluminescent microparticle immunoassay for quantitative determination of dehydroepiandrosterone sulfate

HUANG Mingcong1, ZHANG Xiaoli2, LIN Guanghua2, WENG Zuxing1, SUN Xudong1, GE Shengxiang3, ZHANG Jun3★
(1. Xiamen Innodx Biotech Co., Ltd, Xiamen, Fujian, China, 361022; 2. Clinical Laboratory Department, Fujian Medical University Union Hospital, Fuzhou, Fujian, China, 350001; 3. National Institute of Diagnostics and Vaccine Development in Infectious Disease, Xiamen University，Xiamen, Fujian, China, 361102)

[ABSTRACT]Objective To develop chemiluminescent microparticle immunoassay for quantitative determination of dehydroepiandrosterone sulfate. Methods Mice were immunized by dehydroepiandrosterone sulfate artificial complete antigen.Monoclonal antibodies against dehydroepiandrosterone sulfate were prepared by hybridoma technique.Chemiluminescent microparticle immunoassay for quantitative determination of dehydroepiandrosterone sulfate was developed based on competitive binding method.Results27 monoclonal antibodies were obtained. Chemiluminescent microparticle immunoassay for quantitative determination of dehydroepiandrosterone sulfate prototype kit was established. The coefficient r of testing specimens with reagent from Abbott company was over 0．99. Conclusion Development of domestic chemiluminescent microparticle immunoassay for quantitative determination of dehydroepiandrosterone sulfate reagent was laid foundation by this study.

[KEY WORDS]Dehydroepiandrosterone sulfate; Chemiluminescent microparticle immunoassay; Quantitative determination kit

硫酸脫氫表雄酮（dehydroepiandrosterone sulfate，DHEAs）是脫氫表雄酮（dehydroepiandrosterone，DHEA）的硫酸酯形式，是一種主要由腎上腺皮質(zhì)分泌的甾醇類激素［1－2］。作為雄性激素和雌性激素的前體，DHEAs是人體中含量最豐富的甾醇類物質(zhì)，在人血清中的含量比DHEA高250～500倍，比睪酮高100～500倍，比雌二醇高1 000～10 000倍［3－4］。循環(huán)血液中的DHEAs并不具備雄性和雌性激素的生物學(xué)活性，運(yùn)送到靶組織后經(jīng)脫硫酸化生成DHEA，進(jìn)而轉(zhuǎn)變?yōu)椴煌男坌院痛菩约に鼗衔铮l(fā)揮生物學(xué)功能。……