Liu Changqi, Chhabra Guneet S, Sathe Shridhar K
Department of Nutrition, Food and Exercise Sciences, Florida State University , Tallahassee, Florida 32306, United States.
J Agric Food Chem. 2015 Oct 21;63(41):9139-49. doi: 10.1021/acs.jafc.5b03066. Epub 2015 Oct 7.
A commercially available direct sandwich enzyme-linked immunosorbent assay (ELISA) (BioFront Technologies, Tallahassee, FL, USA) using murine anti-pistachio monoclonal antibodies (mAbs) as capture and detection antibodies was evaluated. The assay was sensitive (limit of detection = 0.09 ± 0.02 ppm full fat pistachio, linear detection range = 0.5-36 ppm, 50% maximum signal concentration = 7.9 ± 0.7 ppm), reproducible (intra- and inter-assay variability < 24% CV), and rapid (post-extraction testing time ∼ 1.5 h). The target antigen was stable and detectable in whole pistachio seeds subjected to autoclaving (121 °C, 15 psi, 15, 30 min), blanching (100 °C, 5, 10 min), frying (191 °C, 1 min), microwaving (500, 1000 W, 3 min), and dry roasting (140 °C, 30 min; 168 °C, 12 min). No cross-reactivity was observed in 156 food matrices, each tested at 100,000 ppm, suggesting the ELISA to be pistachio specific. The pistachio recovery ranges for spiked (10 ppm) and incurred (10-50000 ppm) food matrices were 93.1-125.6% and 35.7-112.2%, respectively. The assay did not register any false-positive or -negative results among the tested commercial and laboratory prepared samples.
对一种市售的直接夹心酶联免疫吸附测定（ELISA）（美国佛罗里达州塔拉哈西市的BioFront Technologies公司）进行了评估，该测定使用鼠抗开心果单克隆抗体（mAb）作为捕获抗体和检测抗体。该测定灵敏（检测限=0.09±0.02 ppm全脂开心果，线性检测范围=0.5 - 36 ppm，最大信号浓度的50% = 7.9±0.7 ppm）、可重复（批内和批间变异<24% CV）且快速（提取后测试时间约1.5小时）。目标抗原在经过高压灭菌（121°C，15 psi，15、30分钟）、热烫（100°C，5、10分钟）、油炸（191°C，1分钟）、微波处理（500、1000 W，3分钟）和干烤（140°C，30分钟；168°C，12分钟）的完整开心果种子中稳定且可检测。在156种食品基质中未观察到交叉反应，每种基质的测试浓度为1,000,000 ppm，这表明该ELISA对开心果具有特异性。加标（10 ppm）和实际污染（10 - 500,000 ppm）食品基质的开心果回收率范围分别为93.1 - 125.6%和35.7 - 112.2%。在测试的商业样品和实验室制备样品中，该测定未出现任何假阳性或假阴性结果。
