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Development of a novel parallel determination platform: a feasibility study tested on a chemiluminescence device
Analytical Methods ( IF 3.1 ) Pub Date : 2017-11-13 00:00:00 , DOI: 10.1039/c7ay02394d Chunjiao Yang 1, 2, 3, 4, 5 , Zhongfeng Sun 6, 7 , Guojun Zhang 1, 2, 3, 4, 5 , Lijuan Wang 1, 2, 3, 4, 5 , Jie Zhang 6, 7 , Xin Zhang 6, 7
Analytical Methods ( IF 3.1 ) Pub Date : 2017-11-13 00:00:00 , DOI: 10.1039/c7ay02394d Chunjiao Yang 1, 2, 3, 4, 5 , Zhongfeng Sun 6, 7 , Guojun Zhang 1, 2, 3, 4, 5 , Lijuan Wang 1, 2, 3, 4, 5 , Jie Zhang 6, 7 , Xin Zhang 6, 7
Affiliation
The potential of obtaining incremental diagnostic information using a parallel assay is attractive. Herein, a novel parallel determination platform was developed for the simultaneous determination of hormones, including human growth hormone (GH), prolactin (PRL), thyroid-stimulating hormone (TSH), follicle-stimulating hormone (FSH) and luteinizing hormone (LH). Polystyrene was used as a support material and the device comprised an antibody capture area, a connecting channel, and an isolating bar. After immobilizing antibodies in the antibody capture area, 50 μL antigens and 1.5 mL of HRP-labelled antibodies were mixed and bound to the capture antibodies by flowing in the connecting channel continually. After adding chemiluminescence substrates, the isolating bar enabled the connecting channel to enter into the detection site in a 1 : 1 ratio. Fluorescein isothiocyanate-labelled bovine serum albumin (FITC-BSA) was used as an internal standard. The developed method was validated in terms of limits of detection (LOD), linearity, recovery and precision. The LOD were 0.01 mIU L−1, 0.01 IU L−1, 0.03 mIU L−1, 0.05 mIU L−1, and 0.01 IU L−1 for GH, FSH, TSH, PRL, and LH, respectively. The assay was linear up to 230 mIU L−1 for GH (r2 = 0.995), 300 IU L−1 for FSH (r2 = 0.991), 100 mIU L−1 for TSH (r2 = 0.997), 8000 mIU L−1 for PRL (r2 = 0.994), and 260 IU L−1 for LH (r2 = 0.992). The mean recoveries were calculated at two concentration levels and the values were found to be between 90.5% and 108.3%. The intra- and inter-assay coefficients of variation were <10% and <11%, respectively. The method was successfully applied to determine serum sample (n = 120) and the results were strongly correlated with the Siemens and Abbott immunoassay, showing a bias offset of 1.7 mIU L−1, 2.4 IU L−1, 0.1 mIU L−1, 7.6 mIU L−1 and 1.5 IU L−1 for GH, FSH, TSH, PRL, and LH. This sensitive and cost-efficient platform is expected to be a powerful tool to measure combinations of various biomarkers.
中文翻译:
新型平行测定平台的开发:在化学发光装置上测试的可行性研究
使用平行测定获得增量诊断信息的潜力很有吸引力。在此,开发了一种新型的平行测定平台,用于同时测定激素,包括人体生长激素(GH),催乳激素(PRL),促甲状腺激素(TSH),促卵泡激素(FSH)和促黄体生成激素(LH) 。聚苯乙烯用作支撑材料,该设备包含抗体捕获区域,连接通道和隔离条。将抗体固定在抗体捕获区域后,将50μL抗原和1.5 mL HRP标记的抗体混合,并通过持续在连接通道中流动而与捕获抗体结合。添加化学发光底物后,隔离条使连接通道以1:1的比例进入检测位点。异硫氰酸荧光素标记的牛血清白蛋白(FITC-BSA)用作内标。所开发的方法在检出限(LOD),线性,回收率和精密度方面得到了验证。LOD为0.01 mIU L-1,0.01 IU大号-1,0.03 MIU大号-1,0.05 MIU大号-1和0.01 IU大号-1为GH,FSH,TSH,PRL,和LH分别。对于GH( r 2 = 0.995),测定高达300mIU L -1 ;对于FSH( r 2 = 0.991),测定高达300 IU L -1 ;对于TSH( r 2 = 0.997),100 mIU L -1(8000 mIU)PRL为L -1( r 2 = 0.994),LH为260 IU L -1( r 2= 0.992)。计算了两个浓度水平下的平均回收率,发现回收率在90.5%和108.3%之间。批内和批间变异系数分别<10%和<11%。该方法被成功地用于测定血清样品(Ñ = 120)并且将结果强烈西门子和雅培免疫相关的,示出了偏置的1.7 MIU错开L -1,2.4 IU大号-1,0.1 MIU大号-1,对于GH,FSH,TSH,PRL和LH,分别为7.6 mIU L -1和1.5 IU L -1。这个灵敏且具有成本效益的平台有望成为衡量各种生物标志物组合的有力工具。
更新日期:2017-11-13
中文翻译:
新型平行测定平台的开发:在化学发光装置上测试的可行性研究
使用平行测定获得增量诊断信息的潜力很有吸引力。在此,开发了一种新型的平行测定平台,用于同时测定激素,包括人体生长激素(GH),催乳激素(PRL),促甲状腺激素(TSH),促卵泡激素(FSH)和促黄体生成激素(LH) 。聚苯乙烯用作支撑材料,该设备包含抗体捕获区域,连接通道和隔离条。将抗体固定在抗体捕获区域后,将50μL抗原和1.5 mL HRP标记的抗体混合,并通过持续在连接通道中流动而与捕获抗体结合。添加化学发光底物后,隔离条使连接通道以1:1的比例进入检测位点。异硫氰酸荧光素标记的牛血清白蛋白(FITC-BSA)用作内标。所开发的方法在检出限(LOD),线性,回收率和精密度方面得到了验证。LOD为0.01 mIU L-1,0.01 IU大号-1,0.03 MIU大号-1,0.05 MIU大号-1和0.01 IU大号-1为GH,FSH,TSH,PRL,和LH分别。对于GH( r 2 = 0.995),测定高达300mIU L -1 ;对于FSH( r 2 = 0.991),测定高达300 IU L -1 ;对于TSH( r 2 = 0.997),100 mIU L -1(8000 mIU)PRL为L -1( r 2 = 0.994),LH为260 IU L -1( r 2= 0.992)。计算了两个浓度水平下的平均回收率,发现回收率在90.5%和108.3%之间。批内和批间变异系数分别<10%和<11%。该方法被成功地用于测定血清样品(Ñ = 120)并且将结果强烈西门子和雅培免疫相关的,示出了偏置的1.7 MIU错开L -1,2.4 IU大号-1,0.1 MIU大号-1,对于GH,FSH,TSH,PRL和LH,分别为7.6 mIU L -1和1.5 IU L -1。这个灵敏且具有成本效益的平台有望成为衡量各种生物标志物组合的有力工具。