Ultra-sensitive One-step Platform for Logic Screening and Detecting of HPV Genotypes via a Programmable Amplification-transcription Cascade

doi:10.1007/s40242-026-5313-z

Chemical Research in Chinese Universities ›› 2026, Vol. 42 ›› Issue (2): 514-525.doi: 10.1007/s40242-026-5313-z

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Ultra-sensitive One-step Platform for Logic Screening and Detecting of HPV Genotypes via a Programmable Amplification-transcription Cascade

TIAN Zilin^1,2, QI Lijuan^1,3, WANG Lingxuan¹, YANG Tianshi^4,5, DU Yan^1,2

1. State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, P. R. China;
2. School of Applied Chemistry and Engineering, University of Science and Technology of China, Hefei 230026, P. R. China;
3. Department of Biochemistry and Biomedical Sciences, McMaster University Hamilton L8S 4K1, Canada;
4. Genetic Diagnosis Center, The First Hospital of Jilin University, Changchun 130012, P. R. China;
5. Medical Laboratory Center, Jilin Province People's Hospital, Changchun 130021, P. R. China

Received:2025-12-31 Online:2026-04-01 Published:2026-04-02
Contact: DU Yan,E-mail:duyan@ciac.ac.cn E-mail:duyan@ciac.ac.cn
Supported by:
This work was supported by the National Natural Science Foundation of China (Nos. 22322410, 22404156 and 22574151), the Major Project of Jilin Province State Key Laboratory, China (No. SKL202402017), and the Major Project of Changchun State Key Laboratory, China (No. 2024GZZ13).

Abstract

Abstract: Ultra-sensitive, versatile, and contamination-resistant nucleic acid detection remains a major challenge in molecular diagnostics, particularly for applications requiring simultaneous pathogen screening and precise genotyping. Herein, we report a programmable, label-free isothermal sensing strategy termed probe-initiated amplification and transcription for high-sensitivity sensing platform (PATSP), which decouples target recognition from signal generation via a universal signal-transduction architecture. In this system, target-specific hybridization events are converted into a universal intermediate trigger that activates a fixed amplification-transcription cascade, generating malachite green (MG) RNA aptamers as the fluorescent output. By integrating loop-mediated isothermal amplification (LAMP) with in situ transcription in a single closed-tube format, PATSP achieves high sensitivity while effectively suppressing aerosol contamination and false-positive signals. Notably, only minimal probe sequence replacement is required for new targets, enabling one-tube multiplex screening using a single primer pair without primer cross-reactivity. Under optimized conditions, PATSP enables attomolar-level detection of multiple human papillomavirus (HPV) genotypes (11, 16, 18, and 52) and human immunodeficiency virus (HIV) over a wide dynamic range. Clinical validation using cervical secretion samples showed 100% concordance with quantitative polymerase chain reaction (qPCR) for both HPV screening and genotyping. Collectively, PATSP provides a robust and broadly applicable platform for nucleic acid diagnostics and point-of-care testing.

Key words: Isothermal amplification, Nucleic acid detection, Human papillomavirus, Genotyping, Fluorescent aptamer

TIAN Zilin, QI Lijuan, WANG Lingxuan, YANG Tianshi, DU Yan. Ultra-sensitive One-step Platform for Logic Screening and Detecting of HPV Genotypes via a Programmable Amplification-transcription Cascade[J]. Chemical Research in Chinese Universities, 2026, 42(2): 514-525.

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Ultra-sensitive One-step Platform for Logic Screening and Detecting of HPV Genotypes via a Programmable Amplification-transcription Cascade

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