College of Forestry, Wildlife and Environment, Forest Products Development Center, Auburn University, Auburn, AL, USA.
Sustainable Bio-Based Lab, Auburn University, Auburn, AL, USA.
J Med Entomol. 2023 May 12;60(3):535-545. doi: 10.1093/jme/tjad008.
Malaria is responsible for over 435,000 deaths annually, mostly occurring in sub-Saharan Africa. Detecting Plasmodium spp. sporozoites (spzs) in the salivary glands of Anopheles (Diptera: Culicidae) vectors with circumsporozoite enzyme-linked immunosorbent assay (csELISA) is an important surveillance method. However, current technological advances are intellectual property and often require of distribution and highly trained users. The transition into paper-based rapid plataforms would allow for decentralization of survillance, especially in areas where it was virtually eliminated. The addition of bio-based materials have shown the potential to improve binding of target antigens, while being widely available. Here, we evaluate the use of chitosan and cellulose nanocrystals (CNC) as antibody carriers and substrate coatings on 96-well plates and on wax hydrophobized paper plates for the detection of Plasmodium falciparum (Pf), P. vivax VK210 (Pv210), and P. vivax VK247 (Pv247). To further improve the user-friendliness of the paper plates a quantitative photograph image-based color analysis was done. Interactions between the materials and the assay antibodies were studied by quartz crystal microbalance with dissipation monitoring (QCM-D). Overall, the addition of chitosan increased the interaction with antibodies and enhanced signaling in all tests. This work demonstrated that the adaptation of a PcsELISA shows potential as a cost-effective alternative assay platform easily adaptable in deployable testing sites that also showed reduction in reagent volumes by 80% and assay run time by seventh. While dipstick assays were previously developed, paper-based assays are a cost-effective and field-deployable alternative, reducing volumes of reagents that could be used in malaria control and elimination settings.
疟疾每年导致超过 43.5 万人死亡，主要发生在撒哈拉以南非洲地区。利用环子孢子蛋白酶联免疫吸附试验（csELISA）检测在按蚊（双翅目：蚊科）唾液腺中的疟原虫 spp. （spzs）孢子是一种重要的监测方法。然而，当前的技术进步涉及知识产权，并且通常需要分布和高度训练有素的用户。向基于纸张的快速平台的转变将允许监测去中心化，特别是在那些实际上已经消除疟疾的地区。生物基材料的加入显示了提高目标抗原结合能力的潜力，同时广泛可用。在这里，我们评估了壳聚糖和纤维素纳米晶体（CNC）作为抗体载体和底物涂层在 96 孔板以及蜡疏水化纸盘上对恶性疟原虫（Pf）、间日疟原虫 VK210（Pv210）和间日疟原虫 VK247（Pv247）的检测效果。为了进一步提高纸盘的易用性，进行了基于定量照片图像的颜色分析。通过石英晶体微天平耗散监测（QCM-D）研究了材料与检测抗体之间的相互作用。总的来说，壳聚糖的添加增加了与抗体的相互作用，并增强了所有测试中的信号。这项工作表明，PcsELISA 的适应具有作为一种具有成本效益的替代检测平台的潜力，易于在可部署的测试点中进行调整，同时还将试剂体积减少了 80%，检测运行时间缩短了七分之二。虽然之前已经开发了侧流试验，但基于纸张的试验是一种具有成本效益且可在现场部署的替代方法，可以减少在疟疾控制和消除环境中使用的试剂量。
