ORIGINAL RESEARCH article
Front. Bioeng. Biotechnol.
Sec. Biosensors and Biomolecular Electronics
This article is part of the Research TopicAdvanced and Robust Point-of-Care Testing Systems: Integrated Emerging Technology and StrategiesView all articles
Multiplex on-chip detection of Aspergillus by integrated ultrasonication-based bead beating lysis and magnetic beads direct amplification
Provisionally accepted
- 1Fudan University, Shanghai, China
- 2Wuxi No 5 People's Hospital, Wuxi, China
- 3IngeDx Technologies Co., Ltd, Suzhou, China, Suzhou, China
- 4Huashan Hospital Fudan University, Shanghai, China
- 5Shanghai Sci-Tech Inno Center for Infection & Immunity, Shanghai, China
- 6The East Hospital Affiliated to Tongji University, Shanghai, China
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Invasive aspergillosis (IA) is a life-threatening disease in immunocompromised individuals, creating an urgent need for rapid, sensitive, and user-friendly methods for early detection of Aspergillus spores in clinical and environmental samples. In this study, we developed a rapid ultrasonication-based beads beating (USBB) method for cell lysis and fungal DNA release, along with an integrated magnetic beads-based direct amplification (MBDA) method utilizing microfluidic chip technology. Our flexible thin-film microfluidic chip was developed to enable immediate contacting with an ultrasonic oscillator for nucleic acid extraction. The chip achieved a lysis efficiency of 85.5% for Aspergillus at 10⁵ spores per sample. Magnetic beads releasing Fe ions at 75–860 ng/mL allowed high-efficiency direct PCR amplification. Complete transfer of the nucleic acid extract into PCR amplification led to a 100-fold improvement in detection sensitivity. The integrated USBB-MBDA system completed the entire workflow from sample-in to result-out within 35 minutes (5 minutes fungal DNA release and 30 minutes TaqMan assay). Using this approach, we achieved simultaneous triplex detection of Aspergillus fumigatus, Aspergillus flavus, and Aspergillus niger, with a sensitivity as low as 10 spores per test. Validation on 37 clinical samples showed complete concordance with MALDI-TOF mass spectrometry (MS). This study establishes an integrated nucleic acid extraction and magnetic bead–enabled direct amplification strategy, providing a versatile approach for the development of automated analytical platforms with broad applicability to in vitro diagnostics.
Keywords: Aspergillus fumigatus, Beads beating, Magnetic beads direct amplification, Thin-film microfluidic chip, Ultrasonication lysis
Received: 26 Dec 2025; Accepted: 03 Feb 2026.
Copyright: © 2026 Zhong, Zhang, Huang, Cheng, Li, Wang, Zhang and Guodong. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence:
Tong Zhang
Sui Guodong
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