Rapid, ultrasensitive, and specific RPA-THz system for pathogenic microorganism detection

Xupeng Fu; Lintao Huang; Ying Chen; Menglu Pi; Lin Ma; Hu Cai; Xuehao Wang; Zhihao Chen; Hang Shi; Wenhui Yang; Fulai Zhang; Yang Zhang; Huili Jiang; Zeming Zhou; Changhe Wang; Rong Huang; Juan Zhang; Donghao Cheng; Li An Wu; Airong Qian; Ye Tian

doi:10.1364/BOE.549870

Rapid, ultrasensitive, and specific RPA-THz system for pathogenic microorganism detection

Xupeng Fu, Lintao Huang, Ying Chen, Menglu Pi, Lin Ma, Hu Cai, Xuehao Wang, Zhihao Chen, Hang Shi, Wenhui Yang, Fulai Zhang, Yang Zhang, Huili Jiang, Zeming Zhou, Changhe Wang, Rong Huang, Juan Zhang, Donghao Cheng, Li An Wu, Airong QianYe Tian

School of life sciences

Research output: Contribution to journal › Article › peer-review

Abstract

Pathogenic microorganisms responsible for infectious diseases pose a significant global threat to human health. Existing detection methods, such as qPCR and ELISA, fail to simultaneously meet the requirements for high sensitivity, high specificity, and rapid detection. This study presents an innovative approach for the rapid, specific, and highly sensitive detection of pathogenic microorganisms, particularly Escherichia coli O157:H7 (E. coli O157:H7) and varicella-zoster virus (VZV), by combining recombinase polymerase amplification (RPA) with terahertz time-domain spectroscopy (THz-TDS). The qualitative and quantitative detection method for pathogenic microorganisms was developed and evaluated. The stable and efficient RPA reaction systems were established to specifically amplify the key conserved genes of these pathogens. Then the RPA products were purified, and enriched with MBs. The absorbance spectra were obtained using THz-TDS technology. The linear range of the RPA-THz for detecting E. coli O157:H7 was 0.55 to 5.5 × 10⁴ pg/mL, while for VZV, it was 0.75 to 7.5 × 10³ pg/mL. The limit of detection (LOD) for bacteria and viruses was 0.226 pg/mL and 0.528 pg/mL, respectively, demonstrating better sensitivity than the qPCR (550 pg/mL and 750 pg/mL, respectively). In addition, the whole amplification and detection process was completed in about 35 minutes. Compared to traditional pathogen detection techniques, the primary advantage of the developed RPA-THz method exhibited high accuracy, good reproducibility, and short detection times, enabling non-ionizing, label-free analysis for rapid detection with high sensitivity and specificity of pathogenic microorganisms. This study provides a theoretical foundation and practical demonstration for the fast and precise detection of pathogenic microorganisms. It establishes a crucial research basis for further development of RPA-THz sensors, advancing technological progress in the field of food safety, medical diagnostics, environmental monitoring, and public health.

Original language	English
Pages (from-to)	949-964
Number of pages	16
Journal	Biomedical Optics Express
Volume	16
Issue number	3
DOIs	https://doi.org/10.1364/BOE.549870
State	Published - 1 Mar 2025

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10.1364/BOE.549870

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Fu, X., Huang, L., Chen, Y., Pi, M., Ma, L., Cai, H., Wang, X., Chen, Z., Shi, H., Yang, W., Zhang, F., Zhang, Y., Jiang, H., Zhou, Z., Wang, C., Huang, R., Zhang, J., Cheng, D., Wu, L. A., ... Tian, Y. (2025). Rapid, ultrasensitive, and specific RPA-THz system for pathogenic microorganism detection. Biomedical Optics Express, 16(3), 949-964. https://doi.org/10.1364/BOE.549870

@article{49518a0248a44f9fab902e316f17d9ce,

title = "Rapid, ultrasensitive, and specific RPA-THz system for pathogenic microorganism detection",

abstract = "Pathogenic microorganisms responsible for infectious diseases pose a significant global threat to human health. Existing detection methods, such as qPCR and ELISA, fail to simultaneously meet the requirements for high sensitivity, high specificity, and rapid detection. This study presents an innovative approach for the rapid, specific, and highly sensitive detection of pathogenic microorganisms, particularly Escherichia coli O157:H7 (E. coli O157:H7) and varicella-zoster virus (VZV), by combining recombinase polymerase amplification (RPA) with terahertz time-domain spectroscopy (THz-TDS). The qualitative and quantitative detection method for pathogenic microorganisms was developed and evaluated. The stable and efficient RPA reaction systems were established to specifically amplify the key conserved genes of these pathogens. Then the RPA products were purified, and enriched with MBs. The absorbance spectra were obtained using THz-TDS technology. The linear range of the RPA-THz for detecting E. coli O157:H7 was 0.55 to 5.5 × 104 pg/mL, while for VZV, it was 0.75 to 7.5 × 103 pg/mL. The limit of detection (LOD) for bacteria and viruses was 0.226 pg/mL and 0.528 pg/mL, respectively, demonstrating better sensitivity than the qPCR (550 pg/mL and 750 pg/mL, respectively). In addition, the whole amplification and detection process was completed in about 35 minutes. Compared to traditional pathogen detection techniques, the primary advantage of the developed RPA-THz method exhibited high accuracy, good reproducibility, and short detection times, enabling non-ionizing, label-free analysis for rapid detection with high sensitivity and specificity of pathogenic microorganisms. This study provides a theoretical foundation and practical demonstration for the fast and precise detection of pathogenic microorganisms. It establishes a crucial research basis for further development of RPA-THz sensors, advancing technological progress in the field of food safety, medical diagnostics, environmental monitoring, and public health.",

author = "Xupeng Fu and Lintao Huang and Ying Chen and Menglu Pi and Lin Ma and Hu Cai and Xuehao Wang and Zhihao Chen and Hang Shi and Wenhui Yang and Fulai Zhang and Yang Zhang and Huili Jiang and Zeming Zhou and Changhe Wang and Rong Huang and Juan Zhang and Donghao Cheng and Wu, {Li An} and Airong Qian and Ye Tian",

note = "Publisher Copyright: {\textcopyright} 2025 Optica Publishing Group under the terms of the Optica Open Access Publishing Agreement.",

year = "2025",

month = mar,

day = "1",

doi = "10.1364/BOE.549870",

language = "英语",

volume = "16",

pages = "949--964",

journal = "Biomedical Optics Express",

issn = "2156-7085",

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number = "3",

}

Fu, X, Huang, L, Chen, Y, Pi, M, Ma, L, Cai, H, Wang, X, Chen, Z, Shi, H, Yang, W, Zhang, F, Zhang, Y, Jiang, H, Zhou, Z, Wang, C, Huang, R, Zhang, J, Cheng, D, Wu, LA, Qian, A & Tian, Y 2025, 'Rapid, ultrasensitive, and specific RPA-THz system for pathogenic microorganism detection', Biomedical Optics Express, vol. 16, no. 3, pp. 949-964. https://doi.org/10.1364/BOE.549870

TY - JOUR

T1 - Rapid, ultrasensitive, and specific RPA-THz system for pathogenic microorganism detection

AU - Fu, Xupeng

AU - Huang, Lintao

AU - Chen, Ying

AU - Pi, Menglu

AU - Ma, Lin

AU - Cai, Hu

AU - Wang, Xuehao

AU - Chen, Zhihao

AU - Shi, Hang

AU - Yang, Wenhui

AU - Zhang, Fulai

AU - Zhang, Yang

AU - Jiang, Huili

AU - Zhou, Zeming

AU - Wang, Changhe

AU - Huang, Rong

AU - Zhang, Juan

AU - Cheng, Donghao

AU - Wu, Li An

AU - Qian, Airong

AU - Tian, Ye

PY - 2025/3/1

Y1 - 2025/3/1

N2 - Pathogenic microorganisms responsible for infectious diseases pose a significant global threat to human health. Existing detection methods, such as qPCR and ELISA, fail to simultaneously meet the requirements for high sensitivity, high specificity, and rapid detection. This study presents an innovative approach for the rapid, specific, and highly sensitive detection of pathogenic microorganisms, particularly Escherichia coli O157:H7 (E. coli O157:H7) and varicella-zoster virus (VZV), by combining recombinase polymerase amplification (RPA) with terahertz time-domain spectroscopy (THz-TDS). The qualitative and quantitative detection method for pathogenic microorganisms was developed and evaluated. The stable and efficient RPA reaction systems were established to specifically amplify the key conserved genes of these pathogens. Then the RPA products were purified, and enriched with MBs. The absorbance spectra were obtained using THz-TDS technology. The linear range of the RPA-THz for detecting E. coli O157:H7 was 0.55 to 5.5 × 104 pg/mL, while for VZV, it was 0.75 to 7.5 × 103 pg/mL. The limit of detection (LOD) for bacteria and viruses was 0.226 pg/mL and 0.528 pg/mL, respectively, demonstrating better sensitivity than the qPCR (550 pg/mL and 750 pg/mL, respectively). In addition, the whole amplification and detection process was completed in about 35 minutes. Compared to traditional pathogen detection techniques, the primary advantage of the developed RPA-THz method exhibited high accuracy, good reproducibility, and short detection times, enabling non-ionizing, label-free analysis for rapid detection with high sensitivity and specificity of pathogenic microorganisms. This study provides a theoretical foundation and practical demonstration for the fast and precise detection of pathogenic microorganisms. It establishes a crucial research basis for further development of RPA-THz sensors, advancing technological progress in the field of food safety, medical diagnostics, environmental monitoring, and public health.

AB - Pathogenic microorganisms responsible for infectious diseases pose a significant global threat to human health. Existing detection methods, such as qPCR and ELISA, fail to simultaneously meet the requirements for high sensitivity, high specificity, and rapid detection. This study presents an innovative approach for the rapid, specific, and highly sensitive detection of pathogenic microorganisms, particularly Escherichia coli O157:H7 (E. coli O157:H7) and varicella-zoster virus (VZV), by combining recombinase polymerase amplification (RPA) with terahertz time-domain spectroscopy (THz-TDS). The qualitative and quantitative detection method for pathogenic microorganisms was developed and evaluated. The stable and efficient RPA reaction systems were established to specifically amplify the key conserved genes of these pathogens. Then the RPA products were purified, and enriched with MBs. The absorbance spectra were obtained using THz-TDS technology. The linear range of the RPA-THz for detecting E. coli O157:H7 was 0.55 to 5.5 × 104 pg/mL, while for VZV, it was 0.75 to 7.5 × 103 pg/mL. The limit of detection (LOD) for bacteria and viruses was 0.226 pg/mL and 0.528 pg/mL, respectively, demonstrating better sensitivity than the qPCR (550 pg/mL and 750 pg/mL, respectively). In addition, the whole amplification and detection process was completed in about 35 minutes. Compared to traditional pathogen detection techniques, the primary advantage of the developed RPA-THz method exhibited high accuracy, good reproducibility, and short detection times, enabling non-ionizing, label-free analysis for rapid detection with high sensitivity and specificity of pathogenic microorganisms. This study provides a theoretical foundation and practical demonstration for the fast and precise detection of pathogenic microorganisms. It establishes a crucial research basis for further development of RPA-THz sensors, advancing technological progress in the field of food safety, medical diagnostics, environmental monitoring, and public health.

UR - http://www.scopus.com/inward/record.url?scp=85219116911&partnerID=8YFLogxK

U2 - 10.1364/BOE.549870

DO - 10.1364/BOE.549870

M3 - 文章

AN - SCOPUS:85219116911

SN - 2156-7085

VL - 16

SP - 949

EP - 964

JO - Biomedical Optics Express

JF - Biomedical Optics Express

IS - 3

ER -

Rapid, ultrasensitive, and specific RPA-THz system for pathogenic microorganism detection

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