1. Rapid, sensitive, and visual detection of pseudorabies virus with an RPA-CRISPR/EsCas13d-based dual-readout portable platform.
- Author
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You, Dong, Xu, Tong, Huang, Bing-Zhou, Wu, Fang, Deng, Li-Shuang, Liu, Zhe-Yan, Ge, Liang-Peng, Liu, Zuo-Hua, Sun, Jing, Zeng, Xiu, Zhou, Yuan-Cheng, Lai, Si-Yuan, Ai, Yan-Ru, Huang, Jian-Bo, Zhu, Ling, and Xu, Zhi-Wen
- Subjects
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AUJESZKY'S disease virus , *RESOURCE-limited settings , *POLYMERASE chain reaction , *SWINE farms , *NUCLEIC acids , *COMMUNICABLE diseases - Abstract
Pseudorabies viruses (PRV) pose a major threat to the global pig industry and public health. Rapid, intuitive, affordable, and accurate diagnostic testing is critical for controlling and eradicating infectious diseases. In this study, a portable detection platform based on RPA-CRISPR/EsCas13d was developed. The platform exhibits high sensitivity (1 copy/μL), good specificity, and no cross-reactivity with common pathogens. The platform uses rapid preamplification technology to provide visualization results (lateral flow assays or visual fluorescence) within 1 h. Fifty pig samples (including tissues, oral fluids, and serum) were tested using this platform and real-time quantitative polymerase chain reaction (qPCR), showing 34.0 % (17 of 50) PRV positivity with the portable CRISPR/EsCas13d dual-readout platform, consistent with the qPCR results. These results highlight the stability, sensitivity, efficiency, and low equipment requirements of the portable platform. Additionally, a novel point-of-care test is being developed for clinical use in remote rural and resource-limited areas, which could be a prospective measure for monitoring the progression of pseudorabies and other infectious diseases worldwide. • We have developed and optimized a highly sensitive and specific nucleic acid detection platform based on cas13d. • The detection platform can detect pathogen nucleic acids within 1 h without any special equipment. • We developed two visual methods for visual detection using Cas13d detection platform. • Our platform provides a novel strategy for epidemiological surveillance and control of PRV. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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