Yu-Zhong Zheng1, Jiang-Tao Chen2, Jian Li3, Xian-Jing Wu2, Jin-Zhou Wen4, Xiang-Zhi Liu5, Li-Yun Lin1, Xue-Yan Liang2,5, Hui-Ying Huang5, Guang-Cai Zha1, Pei-Kui Yang1, Lie-Jun Li6, Tian-Yu Zhong7, Long Liu3, Wei-Jia Cheng3, Xiao-Nan Song3, Min Lin1. 1. School of Food Engineering and Biotechnology, Hanshan Normal University, Chaozhou, China. 2. Department of Medical Laboratory, Huizhou Central People's Hospital, Huizhou, China. 3. Department of Human Parasitology, School of Basic Medical Sciences, Hubei University of Medicine, Shiyan, China. 4. Department of Medical Laboratory, Center for Disease Control and Prevention, Chaozhou, China. 5. Department of Medical Laboratory, Chaozhou People's Hospital, Shantou University Medical College, Chaozhou, China. 6. Department of Research and Development, Chaozhou Hybribio Limited Corporation, Chaozhou, China. 7. Department of Laboratory Medicine, First Affiliated Hospital of Gannan Medical University, Ganzhou, China.
Abstract
Background: The emerging Coronavirus Disease-2019 (COVID-19) has challenged the public health globally. With the increasing requirement of detection for SARS-CoV-2 outside of the laboratory setting, a rapid and precise Point of Care Test (POCT) is urgently needed. Methods: Targeting the nucleocapsid (N) gene of SARS-CoV-2, specific primers, and probes for reverse transcription recombinase-aided amplification coupled with lateral flow dipstick (RT-RAA/LFD) platform were designed. For specificity evaluation, it was tested with human coronaviruses, human influenza A virus, influenza B viruses, respiratory syncytial virus, and hepatitis B virus, respectively. For sensitivity assay, it was estimated by templates of recombinant plasmid and pseudovirus of SARS-CoV-2 RNA. For clinical assessment, 100 clinical samples (13 positive and 87 negatives for SARS-CoV-2) were tested via quantitative reverse transcription PCR (RT-qPCR) and RT-RAA/LFD, respectively. Results: The limit of detection was 1 copies/μl in RT-RAA/LFD assay, which could be conducted within 30 min at 39°C, without any cross-reaction with other human coronaviruses and clinical respiratory pathogens. Compared with RT-qPCR, the established POCT assay offered 100% specificity and 100% sensitivity in the detection of clinical samples. Conclusion: This work provides a convenient POCT tool for rapid screening, diagnosis, and monitoring of suspected patients in SARS-CoV-2 endemic areas.
Background: The emerging Coronavirus Disease-2019 (COVID-19) has challenged the public health globally. With the increasing requirement of detection for SARS-CoV-2 outside of the laboratory setting, a rapid and precise Point of Care Test (POCT) is urgently needed. Methods: Targeting the nucleocapsid (N) gene of SARS-CoV-2, specific primers, and probes for reverse transcription recombinase-aided amplification coupled with lateral flow dipstick (RT-RAA/LFD) platform were designed. For specificity evaluation, it was tested with humancoronaviruses, human influenza A virus, influenza B viruses, respiratory syncytial virus, and hepatitis B virus, respectively. For sensitivity assay, it was estimated by templates of recombinant plasmid and pseudovirus of SARS-CoV-2 RNA. For clinical assessment, 100 clinical samples (13 positive and 87 negatives for SARS-CoV-2) were tested via quantitative reverse transcription PCR (RT-qPCR) and RT-RAA/LFD, respectively. Results: The limit of detection was 1 copies/μl in RT-RAA/LFD assay, which could be conducted within 30 min at 39°C, without any cross-reaction with other humancoronaviruses and clinical respiratory pathogens. Compared with RT-qPCR, the established POCT assay offered 100% specificity and 100% sensitivity in the detection of clinical samples. Conclusion: This work provides a convenient POCT tool for rapid screening, diagnosis, and monitoring of suspected patients in SARS-CoV-2 endemic areas.
Authors: Hong Lin; Song Zhao; Yanhong Liu; Lei Shao; Yuying Ye; Nizhen Jiang; Kun Yang Journal: Front Cell Infect Microbiol Date: 2022-06-24 Impact factor: 6.073
Authors: Yilin Liu; Li Zhan; Jesse W Shen; Bàrbara Baro; Andrea Alemany; James Sackrison; Oriol Mitjà; John C Bischof Journal: ACS Appl Nano Mater Date: 2021-12-14