Advancements in the synergy of isothermal amplification and CRISPR-cas technologies for pathogen detection

Xiaolei Mao; Minghui Xu; Shuyin Luo; Yi Yang; Jiaye Zhong; Jiawei Zhou; Huayan Fan; Xiaoping Li; Zhi Chen

doi:10.3389/fbioe.2023.1273988

Advancements in the synergy of isothermal amplification and CRISPR-cas technologies for pathogen detection

Front Bioeng Biotechnol. 2023 Oct 10:11:1273988. doi: 10.3389/fbioe.2023.1273988. eCollection 2023.

Authors

Xiaolei Mao^#¹, Minghui Xu^#¹, Shuyin Luo¹, Yi Yang¹, Jiaye Zhong¹, Jiawei Zhou¹, Huayan Fan¹, Xiaoping Li^{1

2}, Zhi Chen³

Affiliations

¹ Key Laboratory of Pollution Exposure and Health Intervention of Zhejiang Province, Shulan International Medical College, Zhejiang Shuren University, Hangzhou, China.
² Faculty of Medicine, Macau University of Science and Technology, Macau, China.
³ Shulan International Medical College, Zhejiang Shuren University, Hangzhou, China.

^# Contributed equally.

Abstract

In the realm of pathogen detection, isothermal amplification technology has emerged as a swift, precise, and sensitive alternative to conventional PCR. This paper explores the fundamental principles of recombinase polymerase amplification (RPA) and recombinase-aid amplification (RAA) and reviews the current status of integrating the CRISPR-Cas system with RPA/RAA techniques. Furthermore, this paper explores the confluence of isothermal amplification and CRISPR-Cas technology, providing a comprehensive review and enhancements of existing combined methodologies such as SHERLOCK and DETECTR. We investigate the practical applications of RPA/RAA in conjunction with CRISPR-Cas for pathogen detection, highlighting how this integrated approach significantly advances both research and clinical implementation in the field. This paper aims to provide readers with a concise understanding of the fusion of RPA/RAA and CRISPR-Cas technology, offering insights into their clinical utility, ongoing enhancements, and the promising prospects of this integrated approach in pathogen detection.

Keywords: CRISPR-cas; Cas12; Cas13; DETECTR; rapid inspection technology improvement; recombinase polymerase amplification; recombinase-aid amplification; sherlock.

Publication types

Review

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This study was financially supported by the 2023 National Student Innovation and Entrepreneurship Training Program Project (Item Number: 202311842021X), Analysis and Test Foundation for Scientists, Zhejiang Shuren University and Key Laboratory of Biomarkers and In Vitro Diagnosis Translation of Zhejiang province (KFJJ2023005). The funder did not participate in the designing, performing, or reporting of the current study.