Rapid extended-gate field-effect transistor-type biosensor composed of truncated DNA aptamer and UiO-66 metal organic framework nanoparticle for HPIV detection in Bronchoalveolar Lavage Fluid

Abstract

The human parainfluenza virus (HPIV), the RNA virus similar to SARS-CoV-2, is a leading cause of both upper and lower respiratory diseases, with potentially fatal outcomes. Thus, early diagnosis of HPIV is crucial. However, current HPIV diagnostic methods face certain limitations. Here, we developed a rapid extended-gate field-effect transistor (R-EGFET)-based biosensor consisting of the HPIV aptamer and UiO-66 metal organic framework (MOF) nanoparticle, which is capable of the rapid and specific detection of HPIV. We synthesized an aptamer that selectively detects HPIV hemagglutinin-neuraminidase (HN protein) using systematic evolution of ligands by the exponential enrichment (SELEX) method. Then, the truncation process was achieved to reduce the aptamer production cost. In addition to, UiO-66 MOF nanoparticle is suitable as a biosensor due to its enhanced electrical sensitivity. The R-EGFET was selected as the detection platform for its compact design, low cost, and high sensitivity. In addition, incorporating alternating-current electrothermal flow (ACEF) technology reduced the target binding time to 10 min, enabling rapid detection. Thereby, HPIV was detected based on electrical signal changes across the extended-gated membrane using Au microelectrodes. The limit of detection (LOD) of the HPIV HN protein was 22.254 fM in buffer and 36.202 fM in human serum. In clinically relevant samples, the sensor achieved LODs of 9.961 PFU/mL in artificial saliva and 15.273 PFU/mL in bronchoalveolar lavage fluid (BALF), highlighting its potential for practical diagnostic applications. Owing to its rapid detection and high selectivity, the proposed biosensor is expected to be useful for clinical diagnosis of HPIV.