Department of Chemistry, University of Central Florida, Orlando, Florida 32816, United States.
Anal Chem. 2024 Apr 16;96(15):5752-5756. doi: 10.1021/acs.analchem.3c05962. Epub 2024 Apr 1.
Viruses are the primary cause of many infectious diseases in both humans and animals. Various testing methods require an amplification step of the viral RNA sample before detection, with quantitative reverse transcription polymerase chain reaction (RT-qPCR) being one of the most widely used along with lesser-known methods like Nucleic Acid Sequence-Based Amplification (NASBA). NASBA offers several advantages, such as isothermal amplification and high selectivity for specific sequences, making it an attractive option for low-income facilities. In this research, we employed a single electrochemical biosensor (E-Biosensor) designed for potentially detecting any virus by modifying the NASBA protocol. In this modified protocol, a reverse primer is designed with an additional 22-nucleotide sequence (tag region) at the 5'-end, which is added to the NASBA process. This tag region becomes part of the final amplicon generated by NASBA. It can hybridize with a single specific E-Biosensor probe set, enabling subsequent virus detection. Using this approach, we successfully detected three different viruses with a single E-Biosensor design, demonstrating the platform's potential for virus detection.
病毒是导致人类和动物许多传染病的主要原因。各种检测方法都需要对病毒 RNA 样本进行扩增步骤，其中最广泛使用的方法之一是定量逆转录聚合酶链反应（RT-qPCR），还有一些不太知名的方法，如核酸序列扩增（NASBA）。NASBA 具有许多优势，例如等温扩增和对特定序列的高选择性，使其成为低收入设施的一个有吸引力的选择。在这项研究中，我们采用了一种单一的电化学生物传感器（E-Biosensor），通过修改 NASBA 方案来设计用于潜在检测任何病毒的方案。在这个修改后的方案中，在反向引物的 5'-端设计了一个额外的 22 个核苷酸序列（标签区域），该区域被添加到 NASBA 过程中。这个标签区域成为由 NASBA 生成的最终扩增子的一部分。它可以与单个特定的 E-Biosensor 探针组杂交，从而能够进行后续的病毒检测。通过这种方法，我们使用单个 E-Biosensor 设计成功地检测到了三种不同的病毒，证明了该平台用于病毒检测的潜力。
