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Selective Sensing and Imaging of Penicillium italicum Spores and Hyphae Using Carbohydrate–Lectin Interactions
ACS Sensors ( IF 8.2 ) Pub Date : 2018-02-20 00:00:00 , DOI: 10.1021/acssensors.7b00934 Idris Yazgan , Jing Zhang , Victor Kariuki , Ayfer Akgul 1 , Lauren E. Cronmiller , Ali Akgul 2 , Francis Osonga , Abbey McMahon , Yang Gao , Gaddi Eshun , Seokheun Choi , Omowunmi A. Sadik
ACS Sensors ( IF 8.2 ) Pub Date : 2018-02-20 00:00:00 , DOI: 10.1021/acssensors.7b00934 Idris Yazgan , Jing Zhang , Victor Kariuki , Ayfer Akgul 1 , Lauren E. Cronmiller , Ali Akgul 2 , Francis Osonga , Abbey McMahon , Yang Gao , Gaddi Eshun , Seokheun Choi , Omowunmi A. Sadik
Affiliation
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The blue-green mold Penicillium italicum is among the most problematic post-harvest plant infections limiting the integrity of citrus and many other crops during storage and transportation, but there is no sensor for its on-site or field detection. We hereby, for the first time, report the development of novel biomolecular sensor for assessing the presence of P. italicum spores and hyphae using carbohydrate-lectin recognitions. Two approaches were developed: (i) lateral tests using standalone poly(amic) acid (PAA) membranes and glass surfaces and (ii) quantitative tests on 96-well polystyrene plates and paper electrodes. In both cases, the surfaces were functionalized with novel derivatized sugar based ligands while staining was performed with gold nanoparticles. Both approaches provided strong signals for 104 spores/mL of P. italicum isolated from experimentally infected lemons as the lowest-reliable concentration. The 96-well plate-based gave the most sensitive detection with a 4 × 102 spores/mL limit of detection, a linear dynamic range between 2.9 × 103 and 6.02 × 104 spores/mL (R2 = 0.9939) and standard deviation of less than 5% for five replicate measurements. The selectivity of the ligands was tested against Trichaptum biforme, Glomerulla cingulata (Colletotrichum gloeosporioides), and Aspergillus nidulans fungi species. The highest selectivity was obtained using the sugar-based gold-nanoparticles toward both the spores and the hyphae of P. italicum. The advanced specificity was provided by the utilized sugar ligands employed in the synthesis of gold nanoparticles and was independent from size and shapes of the AuNPs. Accuracy of the sensor response showed dramatic dependence on the sample preparation. In the case of 5–10 min centrifugation at 600 rpm, the spores can be isolated free from hyphae and conidiophore, for which spiked recovery was up to 95% (std ±4). In contrast, for gravity-based precipitation of hyphae, the spiked recovery was 88% (std 11).
中文翻译：
碳水化合物-凝集素相互作用对意大利青霉孢子和菌丝的选择性传感和成像
蓝绿色霉菌意大利青霉菌是收获后问题最严重的植物感染之一，在储存和运输过程中限制了柑橘和许多其他农作物的完整性，但没有用于现场或田间检测的传感器。在此，我们首次报告用于评估意大利疟原虫存在的新型生物分子传感器的开发。孢子和菌丝利用碳水化合物-凝集素识别。开发了两种方法：（i）使用独立的聚（酰胺）酸（PAA）膜和玻璃表面进行横向测试，以及（ii）在96孔聚苯乙烯板和纸电极上进行定量测试。在这两种情况下，表面均用新型衍生糖基配体进行功能化，而金纳米颗粒进行染色。两种方法都为从实验感染的柠檬中分离出的10 4个孢子/意大利Italum的最低可靠浓度提供了强有力的信号。基于96孔板的样品以4×10 2孢子/ mL的检测限进行了最灵敏的检测，线性动态范围介于2.9×10 3和6.02×10 4之间孢子/ mL（R 2 = 0.9939），五次重复测量的标准偏差小于5％。的配位体的选择性为对所测试Trichaptum biforme，Glomerulla炭疽病（炭疽病）和构巢曲霉的真菌物种。使用糖基金纳米颗粒对意大利假单胞菌的孢子和菌丝均获得最高的选择性。先进的特异性是由金纳米颗粒合成中使用的糖配体提供的，并且与AuNP的大小和形状无关。传感器响应的准确性显示出对样品制备的极大依赖。在以600 rpm离心5-10分钟的情况下，可以分离出没有菌丝和分生孢子的孢子，其加标回收率最高为95％（std±4）。相反，对于基于重力的菌丝沉淀，加标回收率为88％（标准11）。
更新日期：2018-02-20
中文翻译：
碳水化合物-凝集素相互作用对意大利青霉孢子和菌丝的选择性传感和成像
蓝绿色霉菌意大利青霉菌是收获后问题最严重的植物感染之一，在储存和运输过程中限制了柑橘和许多其他农作物的完整性，但没有用于现场或田间检测的传感器。在此，我们首次报告用于评估意大利疟原虫存在的新型生物分子传感器的开发。孢子和菌丝利用碳水化合物-凝集素识别。开发了两种方法：（i）使用独立的聚（酰胺）酸（PAA）膜和玻璃表面进行横向测试，以及（ii）在96孔聚苯乙烯板和纸电极上进行定量测试。在这两种情况下，表面均用新型衍生糖基配体进行功能化，而金纳米颗粒进行染色。两种方法都为从实验感染的柠檬中分离出的10 4个孢子/意大利Italum的最低可靠浓度提供了强有力的信号。基于96孔板的样品以4×10 2孢子/ mL的检测限进行了最灵敏的检测，线性动态范围介于2.9×10 3和6.02×10 4之间孢子/ mL（R 2 = 0.9939），五次重复测量的标准偏差小于5％。的配位体的选择性为对所测试Trichaptum biforme，Glomerulla炭疽病（炭疽病）和构巢曲霉的真菌物种。使用糖基金纳米颗粒对意大利假单胞菌的孢子和菌丝均获得最高的选择性。先进的特异性是由金纳米颗粒合成中使用的糖配体提供的，并且与AuNP的大小和形状无关。传感器响应的准确性显示出对样品制备的极大依赖。在以600 rpm离心5-10分钟的情况下，可以分离出没有菌丝和分生孢子的孢子，其加标回收率最高为95％（std±4）。相反，对于基于重力的菌丝沉淀，加标回收率为88％（标准11）。
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