Department of Parasitology, Banha University Faculty of Medicine, Banha, Egypt.
Pathology Department (Microbiology Unit), Jouf University College of Medicine, Sakaka, Saudi Arabia.
Sci Rep. 2020 Mar 30;10(1):2. doi: 10.1038/s41598-019-55492-1.
Covalent loading or directional binding of biomolecules on gold nanoparticles (AuNPs) could lead to better results than simple direct adsorption for an enhanced ELISA application. The use of Mini-Parasep solvent-free (SF) without ether or ethyl acetate for the clean and efficient concentration of protozoa cysts, it is a single-use device for in vitro diagnostic use only. In this work, we used Mini-Parasep SF for the detection of giardia cysts in comparison to direct smear and Merthiolate-Iodine Formaldehyde Concentration (MIFC) technique in addition to its use in antigen detection by AuNPs biomolecule loading using rabbit polyclonal antibodies (pAb) against purified Giardia antigen (PGA). As a result, Mini-Parasep SF was the most effective method for Giardia cyst detection and regarding optimization of Mini-Parasep antigen detection, our data showed increased sensitivity and specificity of nano-sandwich ELISA to 92% and 94% respectively and increased positive predictive value (PPV) and negative predictive value (NPV) to 88.64% and 95.91% respectively. In conclusion, this research provides that Mini-Parasep SF concentrator enhanced Giardia cyst detection and improved antigen preparation for AuNPs sandwich ELISA in giardiasis diagnosis. The advantages of this method are the short assay time and the raised accuracy of antigen detection providing concentrated samples without the risk of solvent use and being a disposable Mini-Parasep it helps in giardia antigen purification as well as raising the sensitivity and specificity of ELISA through binding AuNPs.
金纳米粒子（AuNPs）上生物分子的共价负载或定向结合可能比简单的直接吸附产生更好的结果，从而增强 ELISA 应用。Mini-Parasep 无溶剂（SF）可用于清洁、高效浓缩原生动物孢囊，无需使用乙醚或乙酸乙酯，它是一种仅供体外诊断使用的一次性设备。在这项工作中，我们使用 Mini-Parasep SF 检测贾第虫孢囊，与直接涂片和 Merthiolate-Iodine Formaldehyde Concentration（MIFC）技术进行比较，此外还使用 Mini-Parasep SF 检测 AuNPs 生物分子负载兔多克隆抗体（pAb）对纯化贾第虫抗原（PGA）的抗原检测。结果表明，Mini-Parasep SF 是检测贾第虫孢囊最有效的方法。关于 Mini-Parasep 抗原检测的优化，我们的数据显示纳米三明治 ELISA 的灵敏度和特异性分别提高到 92%和 94%，阳性预测值（PPV）和阴性预测值（NPV）分别提高到 88.64%和 95.91%。总之，这项研究表明，Mini-Parasep SF 浓缩器增强了贾第虫孢囊的检测，并改善了用于贾第虫病诊断的 AuNPs 三明治 ELISA 的抗原制备。该方法的优点是检测时间短，抗原检测准确性提高，提供浓缩样品，无需使用溶剂，且为一次性 Mini-Parasep，有助于贾第虫抗原的纯化，并通过结合 AuNPs 提高 ELISA 的灵敏度和特异性。
