Article
Immunology
Shuang Liu, Siyuan Huang, Fang Li, Yuanyuan Sun, Jin Fu, Fei Xiao, Nan Jia, Xiaolan Huang, Chunrong Sun, Juan Zhou, Yi Wang, Dong Qu
Summary: The study developed a simple, rapid, sensitive, and specific detection platform for P. aeruginosa infection. The newly developed P. aeruginosa-CRISPR-RPA assay showed high sensitivity and specificity, and could complete P. aeruginosa detection within half an hour.
FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY
(2023)
Article
Microbiology
Feina Li, Jing Xiao, Haiming Yang, Yao Yao, Jieqiong Li, Huiwen Zheng, Qian Guo, Xiaotong Wang, Yuying Chen, Yajie Guo, Yonghong Wang, Chen Shen
Summary: In this study, a rapid and efficient method utilizing RPA-CRISPR/Cas12a technology was proposed for the diagnosis of Mycoplasma pneumoniae. This method eliminates the need for expensive instruments and specialized personnel. The entire process, including sample extraction, RPA reaction, CRISPR/Cas12a detection, and visual observation, can be completed within 1 hour with high sensitivity and specificity.
FRONTIERS IN MICROBIOLOGY
(2022)
Article
Chemistry, Analytical
Yiran Xiao, Honglin Ren, Han Wang, Deying Zou, Yixin Liu, Haosong Li, Pan Hu, Yansong Li, Zengshan Liu, Shiying Lu
Summary: A low-cost platform based on the CRISPR/Cas12a system was designed for the rapid detection of Listeria monocytogenes. This method combines CRISPR/Cas12a and recombinase polymerase amplification (RPA) to provide a contamination-free platform for nucleic acid detection with high specificity and ultrasensitivity. The optimized method can specifically detect target DNA in 25 minutes and successfully detect LM in spiked pork samples and natural meat samples.
Article
Microbiology
Hongjian Zhang, Meng Zhao, Siyun Hu, Kairu Ma, Jixu Li, Jing Zhao, Xin Wei, Lina Tong, Shengqiang Li
Summary: In this study, a real-time recombinase polymerase amplification (RPA) assay for pathogenic Y. enterocolitica was developed based on the ail gene. The optimized RPA conditions allowed the detection to be completed within 20 min at an isothermal temperature of 38 degrees C. The assay showed high sensitivity and specificity, making it a valuable tool for the early diagnosis of Y. enterocolitica.
Article
Parasitology
Peipei Cheng, Yuting Wu, Shuangshuang Guo, Xiaoyu Ma, Chenzhong Fei, Feiqun Xue, Chuangang Zhu, Mi Wang, Feng Gu
Summary: In this study, a method combining recombinase polymerase amplification (RPA) with clustered regularly interspaced short palindromic repeats (CRISPR)/Cas12a system was developed to detect Eimeria species in chicken fecal samples. The method showed high specificity and sensitivity, and enabled rapid detection of Eimeria species for on-site testing.
VETERINARY PARASITOLOGY
(2022)
Article
Chemistry, Multidisciplinary
Gaihua Cao, Jiangbo Dong, Xiaolong Chen, Peng Lu, Yifan Xiong, Lan Peng, Jiawei Li, Danqun Huo, Changjun Hou
Summary: In this study, a strategy (MPT-Cas12a/13a) was developed that combines CRISPR/Cas12a and Cas13a for simultaneous detection of CaMV35S and T-nos. This strategy utilizes multiplex PCR and transcription to achieve simultaneous detection with different signals in the same space. The MPT-Cas12a/13a showed excellent sensitivity, with detection limits as low as 11 copies of T-nos and 13 copies of CaMV35S, and demonstrated outstanding specificity and anti-interference ability in actual sample analysis. Therefore, it has the potential to be used for the detection of GM crops.
CHEMICAL COMMUNICATIONS
(2022)
Article
Food Science & Technology
Han Wang, Yu Zheng, Hong-Lin Ren, Yi-Ran Xiao, Cong Wang, Jiang Chang, Yu-Xi Guo, Pan Hu, Yan-Song Li, Zeng-Shan Liu, Shi-Ying Lu
Summary: A lateral flow immunochromatographic strip targeting on virulence gene ail was developed using recombinase polymerase amplification and quantum dot nanobeads for rapid detection of pathogenic Yersinia enterocolitica in meat samples with high sensitivity and accuracy.
LWT-FOOD SCIENCE AND TECHNOLOGY
(2023)
Article
Agronomy
Lingyu Zeng, Sizhu Zheng, Vaclav Stejskal, George Opit, Radek Aulicky, Zhihong Li
Summary: A new and rapid visual detection assay based on RPA and CRISPR/Cas12a system has been developed to differentiate khapra beetle from similar species. This method can be completed within 40 minutes and has high sensitivity for detecting low levels of DNA at room temperature, which helps to prevent the spread of khapra beetle.
PEST MANAGEMENT SCIENCE
(2023)
Article
Agronomy
Kittisak Buddhachat, Nattaporn Sripairoj, Onchira Ritbamrung, Phithak Inthima, Kumrop Ratanasut, Thanita Boonsrangsom, Tepsuda Rungrat, Pongsanat Pongcharoen, Kawee Sujipuli
Summary: A specific and simple method, the recombinase polymerase amplification (RPA) assay assisted with CRISPR-cas12a assay (RAC), was developed for the detection of Xanthomonas oryzae pv. oryzae (Xoo) without the need for DNA extraction. The RAC system using the Xoo4009 locus showed higher specificity compared to the Xoo80 locus, enabling the diagnosis of Xoo presence in both artificially inoculated and naturally infected rice samples. This developed RAC system exhibited potential for early detection of bacterial leaf blight (BLB) disease outbreak in rice fields.
Article
Biochemical Research Methods
Siyuan Liu, Dagang Tao, Yuying Liao, Yalan Yang, Shouzhang Sun, Yulan Zhao, Peng Yang, Yijie Tang, Bin Chen, Yonggang Liu, Shengsong Xie, Zhonglin Tang
Summary: This study developed a new method for highly sensitive and visual detection of PRRSV within 25 minutes by integrating the RT-RPA-Cas12a system with a single stranded DNA-fluorescently quenched reporter. The technique showed single copy sensitivity and no cross-reactivity with other porcine viruses. The results were highly accurate when compared with quantitative RT-qPCR, making it a promising tool for on-site detection of PRRSV.
ACS SYNTHETIC BIOLOGY
(2021)
Article
Chemistry, Analytical
Lianyu Lu, Huimin Zhang, Fanghe Lin, Leiji Zhou, Zhi Zhu, Chaoyong Yang
Summary: The development of new diagnostic platforms for sensitive and rapid detection of bacterial pathogens is significant for improving precision medication for infectious disease and food safety control. The DMF-Cas12a method, which integrates the CRISPR/Cas12a system with a digital microfluidic chip, enables rapid, sensitive, and automated analysis of bacterial pathogens. The platform shows good potential in automated and highly sensitive pathogen detection for a wide variety of biological applications.
SENSORS AND ACTUATORS B-CHEMICAL
(2023)
Article
Veterinary Sciences
Chenglong Li, Nan Lin, Zhihua Feng, Minhua Lin, Biyun Guan, Kunsen Chen, Wangwang Liang, Qiaohuang Wang, Miaomiao Li, Yu You, Qi Chen
Summary: A new molecular assay combining RPA and CRISPR/Cas12a technology was developed to detect the virulence genes causing AHPND in shrimp. The assay showed rapid and specific detection, with the addition of a lateral flow strip readout for visualization. Field tests demonstrated high predictive agreement.
FRONTIERS IN VETERINARY SCIENCE
(2022)
Article
Food Science & Technology
Liyun Lin, Guangcai Zha, Huagui Wei, Yuzhong Zheng, Peikui Yang, Yaqun Liu, Mouquan Liu, Zhonghe Wang, Xianghui Zou, Hui Zhu, Qiulan Luo, JinQuan Li, Min Lin
Summary: The study developed a detection method for Staphylococcus aureus in food using a combination of recombinase polymerase amplification (RPA) and CRISPR-Cas12a technology. The method demonstrated high sensitivity in detecting low quantities of S. aureus and proved feasible for use in both laboratory and field samples. This accurate and portable detection method has great potential in food safety applications.
Article
Biotechnology & Applied Microbiology
Yifan Xiong, Gaihua Cao, Xiaolong Chen, Jun Yang, Meimei Shi, Yu Wang, Fuping Nie, Danqun Huo, Changjun Hou
Summary: The livestock industry has suffered significant economic damage from African swine fever virus (ASFV) and Capripoxvirus (CaPV), highlighting the urgent need for a reliable detection method. In this study, a rapid and ultra-sensitive DNA detection method, combining recombinase polymerase amplification (RPA) and CRISPR/Cas12a, was developed for ASFV and CaPV. The method, called one-pot-RPA-Cas12a (OpRCas) platform, exhibited high amplification efficiency, constant temperature reaction, and strict target selectivity, providing simplified, accurate, and affordable diagnosis for these viruses.
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
(2022)
Article
Forestry
Jing Zhou, Hanqian Dai, Tingting Dai, Tingli Liu
Summary: This study established a detection method for Phytophthora cambivora using a combination of whole-genome screening, recombinase polymerase amplification (RPA), and CRISPR/Cas12a. The RPA-CRISPR/Cas12a assay showed specific detection of P. cambivora isolates and could detect the presence of P. cambivora in apple fruits. Compared to conventional techniques, this method is faster, more cost-effective, and suitable for on-site detection at entry ports.
Article
Biochemical Research Methods
Pan Hu, Song Zhang, Shi-ying Lu, Meng Li, Jiang Chang, Meng-yun Wang, Chang Li, Ke Zhao, Yu-Ting Guan, Yuan-Yuan Zhang, Yan-Song Li, Yu Zhou, Zeng-Shan Liu, Ou Bai, Hong-Lin Ren
BIOMEDICAL CHROMATOGRAPHY
(2018)
Article
Biochemical Research Methods
Yu Zheng, Pan Hu, Honglin Ren, Han Wang, Qi Cao, Qiang Zhao, Hanxiao Li, Hailing Zhang, Zhanxu Liu, Yansong Li, Cong Wang, Zengshan Liu, Shiying Lu
Summary: A visual and convenient detection method, RPA-SYBR Green I, was developed for pathogenic Yersinia enterocolitica, which can accurately detect the bacteria within a short period of time with sensitivity equivalent to that of conventional PCR, showing 100% consistency with conventional PCR in identifying spiked and meat samples.
ANALYTICAL BIOCHEMISTRY
(2021)
Article
Biochemistry & Molecular Biology
Lixiong Peng, Jiang Chang, Xilin Liu, Shiying Lu, Honglin Ren, Xiaoshi Zhou, Zengshan Liu, Pan Hu
Summary: MC1R is considered a marker of poor prognosis in melanoma and may also play a role in colorectal cancer. The study found that decreased MC1R expression was negatively correlated with P53, MLH1, and PMS2 expression, and high MC1R expression was associated with microsatellite instability. MC1R SNPs were also correlated with clinicopathological characteristics of CRC.
CURRENT ISSUES IN MOLECULAR BIOLOGY
(2021)