数字农科院2.0

A CRISPR/Cas12a-based DNAzyme visualization platform for rapid discrimination of Streptococcus suis serotype 2 versus 1/2 and serotype 1 versus 14

文献类型： 外文期刊

作者： Jing Sun;Jieyu Bai;Yuxuan Huang;Paul R. Langford;Yueling Zhang;Gang Li

作者机构：

关键词： cpsK;CRISPR/Cas12a;G4-DNAzyme;SNP;Streptococcus suis

期刊名称： Talanta

ISSN： 0039-9140

年卷期： 2025 年 294 卷

页码：

收录情况： SCIE（2025版） ; ; EI（2025版）

摘要： Streptococcus suis is a major swine pathogen with serotypes 2 and 14 posing zoonotic risks. However, distinguishing serotypes 1/2 from 2 or 1 from 14 remains challenging due to high similarity in their capsule polysaccharide (CPS) loci. Here, we developed a rapid, equipment-free discriminating platform targeting a single nucleotide polymorphism (SNP) at position 483 of the cpsK gene (G in serotypes 2/14 vs. T/C in 1/2/1). The method integrates recombinase polymerase amplification (RPA) with CRISPR/Cas12a and a G-quadruplex-hemin DNAzyme visualization system. RPA enables isothermal amplification, while CRISPR/Cas12a ensures single-nucleotide specificity by cleaving target DNA. Subsequent DNAzyme catalysis converts colorimetric substrates, enabling naked-eye differentiation via distinct color changes (blue for serotypes 1/2/1 vs. colorless for 2/14). This approach achieved a sensitivity of 101-102 copies per reaction and demonstrated 100 % specificity across 29 S. suis serotypes and related strains. Compared to PCR-based or sequencing methods, our platform eliminates reliance on thermocyclers or fluorescence detectors, reducing costs and operational complexity. The entire workflow, completed within 70 min, offers a practical solution for point-of-care testing in resource-limited settings. By enabling rapid, accurate discrimination, this tool will become a complementary tool for resolving ambiguous serotypes and enhances outbreak management in swine populations and mitigates zoonotic transmission.

分类号：

  • 相关文献

[1]Genome-wide identification of allele-specific expression in response to Streptococcus suis 2 infection in two differentially susceptible pig breeds. Wu, Huayu,Gaur, Uma,Peng, Xianwen,Li, Lianghua,Sun, Hua,Song, Zhongxu,Dong, Binke,Li, Mingbo,Mei, Shuqi,Liu, Guisheng,Wu, Huayu,Gaur, Uma,Peng, Xianwen,Li, Lianghua,Sun, Hua,Dong, Binke,Mei, Shuqi,Liu, Guisheng,Mekchay, Supamit,Wimmers, Klaus,Ponsuksili, Siriluck,Li, Kui.

[2]基于ERA-CRISPR/Cas12a-LFD的犀牛特异性检测技术. 王瑛,张冲,蔡一村,王鑫杰,林颖峥,冯之航,潘良文. 2023

[3]基于RPA-CRISPR/Cas12a的MSTN基因编辑猪检测方法的建立及应用. 迟顺顺,吴丹,王楠,王婉洁,聂雨欣,牟玉莲,刘志国,朱振东,李奎. 2025

[4]猪塞内卡病毒A RT-RAA-CRISPR/Cas12a可视化检测方法的建立. 蒋成辉,郭容霞,李小明,常艳燕,王会宝,刘萍,王晶,杨进才,曾巧英. 2025

[5]基于RTCPA-CRISPR/Cas12a的牛病毒性腹泻病毒检测方法的建立及应用. 蒙琦,常亮,杨明,曹映辉,贺泂杰. 2025

[6]Rapid and Specific Detection of Active SARS-CoV-2 With CRISPR/Cas12a. Xinyi Liu,Yanhua Li,Xin Wang,Yifan Song,Lina Wu,Benyuan Yu,Xiaodong Ma,Peixiang Ma,Ming Liu,Xingxu Huang,Xinjie Wang. 2022

[7]One-Pot Assay for Rapid Detection of Benzimidazole Resistance in Venturia carpophila by Combining RPA and CRISPR/Cas12a. Jia Jie Hu,Duo Liu,Min Zheng Cai,Yang Zhou,Wei Xiao Yin,Chao Xi Luo. 2023

[8]Ultrasensitive SARS-CoV-2 diagnosis by CRISPR-based screen-printed carbon electrode. Wu L.,Wang X.,Wu C.,Cao X.,Tang T.,Huang H.,Huang X.. 2022

[9]CRISPR/Cas12a-mediated Enzymatic recombinase amplification for rapid visual quantitative authentication of halal food. Xiaohui Wang,Wenyu Jin,Yao Yang,Huizi Ma,Honghong Liu,Jiawen Lei,Yuhua Wu,Li Zhang. 2023

[10]Development and application of DETECTR-based rapid detection for pathogenic Bacillusanthracis. Xu J.,Bai X.,Zhang X.,Yuan B.,Lin L.,Guo Y.,Cui Y.,Liu J.,Cui H.,Ren X.,Wang J.,Yuan Y.. 2023

[11]CRISPR/Cas12a-based hypochlorous acid and myeloperoxidase biosensors designed on RESET effect. Jia Yi Ma,Bo Liu,Shabib Raza,Hong Xin Jiang,An Na Tang,De Ming Kong. 2023

[12]An ultrasensitive, rapid and portable method for screening oseltamivir-resistant virus based on CRISPR/Cas12a combined with immunochromatographic strips. Hao Zhang,Fayu Yang,Man Yang,Jing Liu,Mi Wang,Chenzhong Fei,Lifang Zhang,Feiqun Xue,Chuangang Zhu,Yingchun Liu,Feng Gu. 2022

[13]A universal CRISPR/Cas12a-powered intelligent point-of-care testing platform for multiple small molecules in the healthcare, environment, and food. Yuan Zhao,Wenqin Wu,Xiaoqian Tang,Qi Zhang,Jin Mao,Li Yu,Peiwu Li,Zhaowei Zhang. 2023

[14]On-Site and Visual Detection of the H5 Subtype Avian Influenza Virus Based on RT-RPA and CRISPR/Cas12a. Xu Zhou,Siwen Wang,Yue Ma,Yongping Jiang,Yanbing Li,Jianzhong Shi,Guohua Deng,Guobin Tian,Huihui Kong,Xiurong Wang. 2024

[15]Conformational transformation of activator strand directly regulates CRISPR/Cas12a system activity for live cell sensing of multiple biomolecules. Qing Nan Li,Dong Xia Wang,Zhi Qi Dai,Shun Li Wu,Gui Mei Han,Sha Lu,Li Na Zhu,Hong Xin Jiang,De Ming Kong. 2024

[16]Analyte-induced hindrance in the RCA-assisted CRISPR/Cas12a system for homogeneous protein assays. Guobin Mao,Qiaoyu Li,Ziying Zhang,Wei Huang,Qian Luo,Junbiao Dai,Weiren Huang,Yingxin Ma. 2024

[17]A novel HCR-CRISPR/Cas12a immunosensor for the sensitive detection of pesticide residues in animal-derived foods. Yuanshang Wang,Pengfei Du,Xiaohong Wang,Jia Wang,Youyou Lu,Feng He,Haijie Wang,Weiting Wang,Yaobo Liu,Shoujing Wang,Jianfang Cao,Yanli Ma,A. M. Abd El-Aty. 2024

[18]Ultra-Sensitive Detection of the SARS-CoV-2 Nucleocapsid Protein via a Clustered Regularly Interspaced Short Palindromic Repeat/Cas12a-Mediated Immunoassay. Yin, Wen,Li, Leyao,Yang, Yang,Yang, Yuxin,Liang, Ruijing,Ma, Lixin,Dai, Junbiao,Mao, Guobin,Ma, Yingxin. 2024

[19]OPERA-Cas12a: A streamlined one-pot system for specific and sensitive on-site detection of nucleic acids. Xiaohui Wang,Qianqian Liang,Zhifei Liu,Qingqing Xie,Jiawen Lei,Yuhua Wu,Guojun Cheng,Li Zhang. 2024

[20]Rapid detection of Mycoplasma hyopneumoniae by recombinase-aided amplification combined with the CRISPR/Cas12a system. Kaili Li,Tingyu Luo,Yu Zhang,Changwen Li,Hongyan Chen,Changyou Xia,Caixia Gao. 2024

作者其他论文 更多>>