Basso Caroline Rodrigues, Yamakawa Ana Carolina, Cruz Taís Fukuta, Pedrosa Valber Albuquerque, Magro Massimiliano, Vianello Fabio, Araújo Júnior João Pessoa
Biotechnology Institute, São Paulo State University, Botucatu 18607-440, SP, Brazil.
Chemical and Biological Sciences Department, Bioscience Institute, São Paulo State University, Botucatu 18618-000, SP, Brazil.
Pathogens. 2022 May 12;11(5):570. doi: 10.3390/pathogens11050570.
The aim of the current study is to present a low-cost and easy-to-interpret colorimetric kit used to diagnose porcine circovirus 2 (PCV-2) to the naked eye, without any specific equipment. The aforementioned kit used as base hybrid nanoparticles resulting from the merge of surface active maghemite nanoparticles and gold nanoparticles, based on the deposition of specific PCV-2 antibodies on their surface through covalent bonds. In total, 10 negative and 40 positive samples (≥10 DNA copies/µL of serum) confirmed by qPCR technique were tested. PCV-1 virus, adenovirus, and parvovirus samples were tested as interferents to rule out likely false-positive results. Positive samples showed purple color when they were added to the complex, whereas negative samples showed red color; they were visible to the naked eye. The entire color-change process took place approximately 1 min after the analyzed samples were added to the complex. They were tested at different dilutions, namely pure, 1:10, 1:100, 1:1000, and 1:10,000. Localized surface plasmon resonance (LSPR) and transmission electron microscopy (TEM) images were generated to validate the experiment. This new real-time PCV-2 diagnostic methodology emerged as simple and economic alternative to traditional tests since the final price of the kit is USD 4.00.
本研究的目的是展示一种低成本且易于解读的比色试剂盒，用于在无需任何特殊设备的情况下，仅凭肉眼诊断猪圆环病毒2型（PCV - 2）。上述试剂盒以表面活性磁赤铁矿纳米颗粒与金纳米颗粒融合产生的复合纳米颗粒为基础，通过共价键将特定的PCV - 2抗体沉积在其表面。总共测试了10份阴性样本和40份经qPCR技术确认的阳性样本（血清中DNA拷贝数≥10个/微升）。对PCV - 1病毒、腺病毒和细小病毒样本作为干扰物进行了测试，以排除可能的假阳性结果。阳性样本加入复合物后呈现紫色，而阴性样本呈现红色，肉眼可见。在将分析样本加入复合物后，整个颜色变化过程大约在1分钟内发生。对它们在不同稀释度下进行了测试，即原液、1:10、1:100、1:1000和1:10000。生成了局部表面等离子体共振（LSPR）和透射电子显微镜（TEM）图像以验证实验。这种新的实时PCV - 2诊断方法是一种简单且经济的传统检测替代方法，因为该试剂盒的最终价格为4.00美元。
