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Efficient detection of Streptococcus pyogenes based on recombinase polymerase amplification and lateral flow strip

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European Journal of Clinical Microbiology & Infectious Diseases Aims and scope Submit manuscript

Abstract

Purpose

This article aims to establish a rapid visual method for the detection of Streptococcus pyogenes (GAS) based on recombinase polymerase amplification (RPA) and lateral flow strip (LFS).

Methods

Utilizing speB of GAS as a template, RPA primers were designed, and basic RPA reactions were performed. To reduce the formation of primer dimers, base mismatch was introduced into primers. The probe was designed according to the forward primer, and the RPA-LFS system was established. According to the color results of the reaction system, the optimum reaction temperature and time were determined. Thirteen common clinical standard strains and 14 clinical samples of GAS were used to detect the selectivity of this method. The detection limit of this method was detected by using tenfold gradient dilution of GAS genome as template. One hundred fifty-six clinical samples were collected and compared with qPCR method and culture method. Kappa index and clinical application evaluation of the RPA-LFS were carried out.

Results

The enhanced RPA-LFS method demonstrates the ability to complete the amplification process within 6 min at 33 °C. This method exhibits a high analytic sensitivity, with the lowest detection limit of 0.908 ng, and does not exhibit cross-reaction with other pathogenic bacteria.

Conclusions

The utilization of RPA and LFS allows for efficient and rapid testing of GAS, thereby serving as a valuable method for point-of-care testing.

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Data availability

The datasets generated and/or analyzed during the current study are available from the corresponding authors on reasonable request.

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Funding

The project was supported by the Science Foundation of Kangda College of Nanjing Medical University (grant number KD2023KYJJ061), the Health and Wellness Surface Science and Technology Project of Lianyungang City (grant number 202217), the Science and Technology Bureau Key R&D Program (Social Development) Project of Lianyungang City (grant number SF2224), the “521 Project” scientific research funding project of Lianyungang City (grant number LYG06521202157), and the 2023 Jiangsu Province Preventive Medicine Research Project (grant number Yl2023028).

Author information

Author notes

  1. Xu-Zhu Gao and Yu-Die Cao contributed equally to this work.

Authors and Affiliations

  1. Department of Central Laboratory, Lianyungang Hospital Affiliated to Kangda College of Nanjing Medical University, Lianyungang, China

    Xu-Zhu Gao, Yu-Zhi Gao & Tuo Ji

  2. Department of Central Laboratory, Lianyungang Hospital Affiliated to Xuzhou Medical University, Lianyungang, China

    Xu-Zhu Gao, Yu-Zhi Gao & Tuo Ji

  3. Institute of Clinical Oncology, The Second People’s Hospital of Lianyungang City (Cancer Hospital of Lianyungang), Lianyungang, China

    Xu-Zhu Gao, Yu-Zhi Gao & Tuo Ji

  4. The Second People’s Hospital of Lianyungang Affiliated to Bengbu Medical College, Lianyungang, China

    Xu-Zhu Gao, Yu-Die Cao & Tuo Ji

  5. Department of Medicine Laboratory, The Second People’s Hospital of Lianyungang City (Cancer Hospital of Lianyungang), 161 Xingfu Road, Lianyungang, China

    Xu-Zhu Gao, Juan Hu & Tuo Ji

Authors
  1. Xu-Zhu Gao
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  2. Yu-Die Cao
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  3. Yu-Zhi Gao
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  4. Juan Hu
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  5. Tuo Ji
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Contributions

Data curation: Yuzhi Gao; funding acquisition: XuZhu Gao and Tuo Ji; methodology: XuZhu Gao and YuDie Cao; project administration: XuZhu Gao; resources: Juan Hu; writing—original draft: XuZhu Gao; writing—review and editing: Tuo Ji.

Corresponding authors

Correspondence to Juan Hu or Tuo Ji.

Ethics declarations

Ethical approval

The study was approved by the Ethics Committee of the Second People’s Hospital of Lianyungang (protocol code 2023K021). We confirm that all experimental protocols were approved by the Institutional Review Board (or Ethics Committee) of the Second People’s Hospital of Lianyungang. We confirm that all methods were carried out in accordance with relevant guidelines and regulations.

Consent to participate

Informed consent was obtained from all the individuals included in this study.

Competing interests

The authors declare no competing interests.

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Highlights

1. Detection of Streptococcus pyogenes using a combination of recombinase polymerase amplification and lateral flow strips.

2. The combined specific probe and primer exhibited high selectivity and analytic sensitivity for the detection of Streptococcus pyogenes.

3. Simple processing for the rapid (6 min) diagnosis of Streptococcus pyogenes infection.

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Gao, XZ., Cao, YD., Gao, YZ. et al. Efficient detection of Streptococcus pyogenes based on recombinase polymerase amplification and lateral flow strip. Eur J Clin Microbiol Infect Dis 43, 735–745 (2024). https://doi.org/10.1007/s10096-024-04780-4

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  • DOI: https://doi.org/10.1007/s10096-024-04780-4

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