# Ultrasensitive and Multiplexed Target Detection Strategy Based on Photocleavable Mass Tags and Mass Signal Amplification

## TL;DR

This paper introduces a highly sensitive and multiplexed detection system using mass tags and signal amplification for identifying multiple viruses in complex biological samples.

## Contribution

A novel mass spectrometry-based strategy using photocleavable mass tags and signal amplification for ultrasensitive multiplex detection of viral targets.

## Key findings

- The system achieved a detection limit of 415.12 amol for the hepatitis B virus target.
- It successfully detected multiple viruses (HBV, HIV, HCV) simultaneously in mixed ratios with high accuracy.
- The method demonstrated reliable performance in fetal bovine serum under low-abundance conditions.

## Abstract

Co-infections pose significant challenges not only clinically, but also in terms of simultaneous diagnoses. The development of sensitive, multiplexed analytical platforms is critical for accurately detecting viral co-infections, particularly in complex biological environments. In this study, we present a mass spectrometry (MS)-based detection strategy employing a target-triggered hybridization chain reaction (HCR) to amplify signals and in situ photocleavable mass tags (PMTs) for the simultaneous detection of multiple targets. Hairpin DNAs modified with PMTs and immobilized loop structures on magnetic particles (Loop@MPs) were engineered for each target, and their hybridization and amplification efficiency was validated using native polyacrylamide gel electrophoresis (PAGE) and laser desorption/ionization MS (LDI-MS), with silica@gold core–shell hybrid (SiAu) nanoparticles being employed as an internal standard to ensure quantitative reliability. The system exhibited excellent sensitivity, with a detection limit of 415.12 amol for the hepatitis B virus (HBV) target and a dynamic range spanning from 1 fmol to 100 pmol. Quantitative analysis in fetal bovine serum confirmed high accuracy and precision, even under low-abundance conditions. Moreover, the system successfully and simultaneously detected multiple targets, i.e., HBV, human immunodeficiency virus (HIV), and hepatitis C virus (HCV), mixed in various ratios, demonstrating clear PMT signals for each. These findings establish our approach as a robust and reliable platform for ultrasensitive multiplexed detection, with strong potential for clinical and biomedical research.