Infectious and Tropical Disease Research Center, Tabriz University of Medical Sciences, Tabriz 51666-14766, Iran.
Student Research Committee, Department of Parasitology and Mycology, Tabriz University of Medical Sciences, Tabriz 51666-14766, Iran.
Biosensors (Basel). 2021 Apr 25;11(5):134. doi: 10.3390/bios11050134.
Human echinococcosis is a serious parasitic diseasethat still affects millions of people in many parts of the world. Since it can offer a critical threat to people's health, it is important to discover a rapid, convenient, and economical method for detection. Herein, we propose a novel point of care assay, namely, an enhanced immuno-dot-blot assay for diagnosis of cystic echinococcosis (hydatidosis). This method is based on the formation of a sandwich complex between a goldnanoprobe (chitosan-gold nanoparticleprotein A) and hydatid cyst antigen (Ag B), which holds anti-Ag B antibodies. Briefly, protein A was conjugated to chitosan-gold nanoparticles via glutaraldehyde chemistry. Then, Ag B was immobilized on the surface of a nitrocellulose membrane, which was followed by the addition of the sera sample and gold nanoprobes. The positive signals were easily detectable by naked eye. The signal intensity of this biosensor was proportional to the concentration of active anti- antibodies on the surface of the nanoparticles, titer of antibodies in the sera samples, and concentration of Ag B coated on the nitrocellulose membrane. The minimum concentration to use the protein A for conjugation to detect titer of anti- IgGand the concentration of Ag B coated in nitrocellulose membrane were 0.5 and 0.3 mg/mL, respectively. This enhanced immuno-dot-blot assay offers a simple diagnostic technique withoutthe need for expensive equipment for diagnosis of echinococcosis.
人棘球蚴病是一种严重的寄生虫病，仍在世界许多地区影响着数百万人。由于它可能对人们的健康构成严重威胁，因此发现一种快速、方便、经济的检测方法非常重要。在这里，我们提出了一种新的即时检测方法，即增强型免疫斑点印迹法用于诊断囊性棘球蚴病（包虫病）。该方法基于金纳米探针（壳聚糖-金纳米颗粒-蛋白 A）与包虫囊抗原（Ag B）之间形成三明治复合物，该复合物结合了抗 Ag B 抗体。简而言之，蛋白 A 通过戊二醛化学与壳聚糖-金纳米颗粒偶联。然后，Ag B 固定在硝酸纤维素膜的表面，随后加入血清样本和金纳米探针。阳性信号可以通过肉眼轻松检测到。该生物传感器的信号强度与纳米颗粒表面上活性抗抗体的浓度、血清样本中的抗体效价以及涂覆在硝酸纤维素膜上的 Ag B 的浓度成正比。用于检测 IgG 效价的蛋白 A 偶联的最小浓度和涂覆在硝酸纤维素膜上的 Ag B 的浓度分别为 0.5 和 0.3mg/mL。这种增强型免疫斑点印迹法提供了一种简单的诊断技术，无需昂贵的设备即可诊断棘球蚴病。
