机构：[1]State Key Laboratory of Food Nutrition and Safety, Key Laboratory of Industrial Microbiology, Ministry of Education, Tianjin Key Laboratory of Industry Microbiology, National and Local United Engineering Lab of Metabolic Control Fermentation Technology, China International Science and Technology Cooperation Base of Food Nutrition/Safety and Medicinal Chemistry, College of Biotechnology, Tianjin University of Science & Technology, Tianjin, China[2]Department of Clinical Laboratory, Xiangyang Central Hospital, AffiliatedHospital of Hubei University of Arts and Science, Xiangyang, Hubei Province, China[3]Department of Clinical Laboratory, Xiangyang No.1 People's Hospital, Hubei Universitly of Medicine, Xiangyang, China[4]Dynamiker Sub-Center of Beijing Key Laboratory for Mechanisms Research and Precision Diagnosis of Invasive Fungal Disease, Tianjin, China[5]Academy of National Food and Strategic Reserves Administration, Beijing, China[6]Institute of Pediatrics, Faculty of Pediatrics, The Seventh Medical Center of Chinese PLA General Hospital, Beijing, China[7]Department of Pathogen Biology, School of Basic Medical Sciences, Tianjin Medical University, Tianjin, China
Persistent infection with high-risk human papillomavirus (HR-HPV) is the principal etiological factor of cervical cancer. Considering the gradual progression of cervical cancer, the early, rapid, sensitive, and specific identification of HPV, particularly HR-HPV types, is crucial in halting the advancement of the illness. Here, we established a rapid, highly sensitive, and specific HR-HPV detection platform, leveraging the CRISPR/Cas12a assay in conjunction with multienzyme isothermal rapid amplification. Our platform enables the detection and genotyping of 14 types of HR-HPV by using type-specific crRNAs. The outcomes of the detection can be interpreted either through a fluorescence reader or visually. Furthermore, we achieved one-tube multiplex detection of 14 HR-HPV types through the use of multiple amplifications and a crRNA pool. The detection sensitivity of this method is 2 copies/μL with no cross-reactivity, and the results can be obtained within 30 minutes. This method exhibited 100% clinical sensitivity and 100% clinical specificity when applied to 258 clinical specimens. Based on these findings, our CRISPR/Cas-based HR-HPV detection platform holds promise as a novel clinical detection tool, offering a visually intuitive and expedited alternative to existing HPV infection diagnostics and providing fresh perspectives for clinical cervical cancer screening.IMPORTANCEThis study developed a novel high-risk human papillomavirus (HR-HPV) detection platform based on CRISPR/Cas12a technology. This platform not only enables the rapid, highly sensitive, and specific detection and genotyping of 14 types of HR-HPV but also achieves single-tube multiplex detection of 14 HR-HPV types through ingenious design. The outcomes of the detection can be interpreted either through a fluorescence reader or visually. To the best of our knowledge, this is the first paper to utilize CRISPR/Cas diagnostic technology for the simultaneous detection of 14 types of HPV and to evaluate its feasibility in clinical sample detection using a large number of clinical samples. We hope that this work will facilitate the rapid and accurate detection of HPV and promote the broader application of CRISPR/Cas diagnostic technology.
基金：
This research was supported by the National Natural Science Foundation of China (32302218, 82002192), Tianjin Science and Technology Planning Project (24YDTPJC00800), Tianjin Municipal Science and Technology Committee (21JCQNJC01410), China Postdoctoral Science Foundation (2021M702460), Liquor Making Biological Technology and Application of Key Laboratory of Sichuan Province (NJ2023-03), Project Program of Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, and Tianjin Key Laboratory of Industrial Microbiology, China (2022KF0203), General Project of Natural Science Foundation of Hubei Province (2022CFB539, 2022CFD107), Young and middle-aged Talents Project of Hubei Provincial Education Department (Q20222605), ScientificResearch Ability Cultivation Fund of Hubei University of Arts and Science (2021kpgj06), Science and Technology Plan (in the fieldof medical and health care) of Xiangyang (2022YL05B, 2022YL12A, 2022YL07B) and Tianjin graduate research innovation project (2022SKYZ101, 2022SKYZ100).
语种：
外文
PubmedID：
中科院(CAS)分区：
出版当年[2025]版：
大类|2 区生物学
小类|3 区微生物学
最新[2025]版：
大类|2 区生物学
小类|3 区微生物学
第一作者：
第一作者机构：[1]State Key Laboratory of Food Nutrition and Safety, Key Laboratory of Industrial Microbiology, Ministry of Education, Tianjin Key Laboratory of Industry Microbiology, National and Local United Engineering Lab of Metabolic Control Fermentation Technology, China International Science and Technology Cooperation Base of Food Nutrition/Safety and Medicinal Chemistry, College of Biotechnology, Tianjin University of Science & Technology, Tianjin, China
共同第一作者：
通讯作者：
推荐引用方式(GB/T 7714)：
Yin Lijuan,Zhao Ziqian,Wang Chunhua,et al.Development and evaluation of a CRISPR/Cas12a-based diagnostic test for rapid detection and genotyping of HR-HPV in clinical specimens[J].Microbiology Spectrum.2025,13(1):e0225324.doi:10.1128/spectrum.02253-24.
APA：
Yin Lijuan,Zhao Ziqian,Wang Chunhua,Zhou Caihong,Wu Xiuzhen...&Shen Liang.(2025).Development and evaluation of a CRISPR/Cas12a-based diagnostic test for rapid detection and genotyping of HR-HPV in clinical specimens.Microbiology Spectrum,13,(1)
MLA：
Yin Lijuan,et al."Development and evaluation of a CRISPR/Cas12a-based diagnostic test for rapid detection and genotyping of HR-HPV in clinical specimens".Microbiology Spectrum 13..1(2025):e0225324
