# Novel Respiratory Disease Diagnosis Tool: Development of an Au‐ReS2‐Functionalized Extended‐gate Field‐Effect Transistor‐Type Aptasensor for Simultaneous Detection of Granzyme B and Perforin

## TL;DR

A new biosensor detects immune markers in respiratory diseases, offering a faster and more reliable alternative to spirometry.

## Contribution

Development of a dual-biomarker aptasensor using Au-ReS2 and EG-FET for rapid detection of granzyme B and perforin in human serum.

## Key findings

- The biosensor detects granzyme B and perforin at femtomolar levels in human serum within 10 minutes.
- The device shows strong correlation with bronchial conditions in COPD patients.
- The system offers rapid, selective, and sensitive detection, potentially replacing conventional spirometry.

## Abstract

Spirometry is influenced by the patient's subjective condition, which limits the reproducibility of diagnostic results despite being a key diagnostic tool for respiratory diseases. To overcome this, an extended‐gate field‐effect transistor‐type aptasensor for detecting granzyme B (GzmB) and perforin (PRF) is introduced as a proof‐of‐concept for diagnosing localized immune responses in respiratory diseases. The novel GzmB and PRF aptamers are synthesized using systematic evolution of ligands by exponential enrichment and are subsequently truncated to enhance the target‐binding affinity. Au‐ReS2 and the alternating current electrothermal flow technique are applied to amplify the biosensing signal and accelerate detection within 10 min, respectively. Under the 10% human serum, a linear response is observed depending on the target concentration, with the detection limits of 330 fM for GzmB and 440 fM for PRF. The targeted dual‐biomarker indicates a strong clinical correlation with bronchial conditions in chronic obstructive pulmonary disease patients. The proposed device demonstrates clear advantages in rapid, selective, and sensitive detection, suggesting its use as a preemptive diagnostic tool for respiratory diseases. This approach is expected to establish promising diagnostic strategies for early detection and therapeutic monitoring of various respiratory diseases, potentially replacing conventional spirometry.

The simultaneous GzmB and PRF‐targeting EG‐FET‐type biosensor is developed for the diagnosis of respiratory diseases. The biosensor achieves femtomolar‐level detection of targets in human serum and reflects bronchial function according to cytotoxic immune activity in COPD patients. Therefore, the introduced system is proposed as an accurate, rapid, and sensitive diagnostic tool for respiratory diseases, capable of replacing conventional spirometry.© 2026 WILEY‐VCH GmbH