通过肉眼直接观察吸附在表面印迹聚合物(SIP)上的荧光染色微生物是非常具有挑战性的,并且受速度的限制。该方法的准确性和半定量性质。在这项研究中,我们测试了尖孢镰刀菌孢子的存在。cubense种族4(Foc4),导致严重的香蕉枯萎病和减少香蕉植物的面积。这种孢子可以在土壤中休眠,这意味着土壤中分泌分子的检测(分子印迹)可能不准确;聚合酶链反应(PCR)和拉曼光谱等检测方法更准确,但耗时且不便。因此,提出了一种半定量快速检测Foc4SIP的方法。基于ITO导电层,制备可重复使用且肉眼可检测的Foc4-PDMSSIP膜,其位置密度为约9000mm-2。吸附实验表明,当Foc4孢子浓度在104~107CFU/mL之间时,Foc4孢子的吸附数量和荧光强度与浓度密切相关,荧光染色后肉眼可以完全区分。对其他微生物的吸附测试表明,SIP膜完全只识别Foc系列。所有结果与荧光染色后的肉眼观察结果高度一致,Foc4感染土壤实验的结果也接近理想情况。一起来看,这些结果表明,Foc4-PDMSSIP具有快速和半定量检测土壤中Foc浓度的能力,可以为香蕉种植提供良好的支持。该方法在其他真菌疾病的检测中也具有潜在的应用价值。
Direct observation by the naked eye of fluorescence-stained microbes adsorbed on surface imprinted polymers (SIPs) is highly challenging and limited by speed, accuracy and the semiquantitative nature of the method. In this study, we tested for the presence of spores of Fusarium oxysporum f. sp. cubense race 4 (Foc4), which cause severe banana Fusarium wilt disease and reduces the area of banana plants. This kind of spore can become dormant in soil, which means that the detection of secreted molecules (molecular imprinting) in soil may be inaccurate; detection methods such as polymerase chain reaction (PCR) and Raman spectroscopy are more accurate but time-consuming and inconvenient. Therefore, a semiquantitative and rapid SIP detection method for Foc4 was proposed. Based on the ITO conductive layer, a reusable and naked-eye-detectable Foc4-PDMS SIP film was prepared with a site density of approximately 9000 mm-2. Adsorption experiments showed that when the Foc4 spore concentration was between 104 to 107 CFU/mL, the number of Foc4 spores adsorbed and the fluorescence intensity were strongly correlated with the concentration and could be fully distinguished by the naked eye after fluorescence staining. Adsorption tests on other microbes showed that the SIP film completely recognized only the Foc series. All the results were highly consistent with the naked-eye observations after fluorescence staining, and the results of the Foc4-infected soil experiment were also close to the ideal situation. Taken together, these results showed that Foc4-PDMS SIPs have the ability to rapidly and semiquantitatively detect the concentration of Foc in soil, which can provide good support for banana cultivation. This method also has potential applications in the detection of other fungal diseases.