Conneely Gráinne, Aherne Margaret, Lu Huihui, Guilbault George G
Sensors Development Group, Department of Chemistry, University College Cork, Ireland.
Anal Chim Acta. 2007 Jan 30;583(1):153-60. doi: 10.1016/j.aca.2006.09.062. Epub 2006 Oct 6.
In the presented work, a disposable immunosensor for the detection of testosterone, an endogenous steroid hormone, in bovine urine has been developed using screen-printed electrodes (SPEs). Due to concerns over the use of steroid hormones as growth promoters, the EU prohibits their use in food producing animals. Consequently, rigorous screening procedures have been implemented in all member states to detect the illegal administration of such compounds. Competitive immunoassays were developed, initially by enzyme linked immunosorbent assay (ELISA), and subsequently transferred to an electrochemical immunosensor format using disposable screen-printed carbon electrodes. Horseradish peroxidase (HRP) was the enzyme label of choice and chronoamperometric detection was carried out using a tetramethylbenzidine/hydrogen peroxide (TMB/H2O2) substrate system, at +100 mV. The EC50 values obtained for the assay in buffer and urine gave relatively comparable results, 710 pg mL(-1) and 960 pg mL(-1), respectively. The linear range obtained for the assay in buffer extended from 0.03 ng mL(-1) to 40 ng mL(-1); while that in urine ranged from 0.03 ng mL(-1) to 1.6 ng mL(-1). The corresponding limits of detection (LOD) in buffer and urine were 26 pg mL(-1) and 1.8 pg mL(-1). Cross reactivity profiles of the antibody have been examined, with notable cross reactivities with 19-nortestosterone (11.6%) and boldenone (9.86%). Precision studies for the sensor demonstrated adequate reproducibility (CV<13%, n=3) and repeatability (CV<9%, n=3). Recovery data obtained showed good agreement between spiking studies and known concentrations of analyte. Sensors showed stability for 4 days at +4 degrees C. A sensitive, highly specific, inexpensive, disposable immunosensor, showing excellent overall performance for the detection of testosterone in bovine urine, has been developed.
在本研究中，已使用丝网印刷电极（SPEs）开发出一种用于检测牛尿中内源性甾体激素睾酮的一次性免疫传感器。由于担心使用甾体激素作为生长促进剂，欧盟禁止在食用动物中使用此类激素。因此，所有成员国都实施了严格的筛查程序，以检测此类化合物的非法施用情况。竞争性免疫测定法最初是通过酶联免疫吸附测定（ELISA）开发的，随后转移到使用一次性丝网印刷碳电极的电化学免疫传感器形式。辣根过氧化物酶（HRP）是首选的酶标记物，并使用四甲基联苯胺/过氧化氢（TMB/H₂O₂）底物系统在+100 mV下进行计时电流检测。该测定法在缓冲液和尿液中获得的半数有效浓度（EC₅₀）值给出了相对可比的结果，分别为710 pg mL⁻¹和960 pg mL⁻¹。该测定法在缓冲液中的线性范围从0.03 ng mL⁻¹扩展到40 ng mL⁻¹；而在尿液中的线性范围为0.03 ng mL⁻¹至1.6 ng mL⁻¹。缓冲液和尿液中相应的检测限（LOD）分别为26 pg mL⁻¹和1.8 pg mL⁻¹。已检查了抗体的交叉反应谱，与19-去甲睾酮（11.6%）和勃地酮（9.86%）有明显的交叉反应。该传感器的精密度研究表明具有足够的重现性（CV<13%，n = 3）和重复性（CV<9%，n = 3）。获得的回收率数据表明加标研究与已知分析物浓度之间具有良好的一致性。传感器在+4℃下可稳定4天。已开发出一种灵敏、高度特异、廉价的一次性免疫传感器，在检测牛尿中的睾酮方面显示出优异的整体性能。
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