Specific and visual detection of EBOV based on a one-pot RT-RAA-CRISPR/Cas12a assay

Zanheng Huang; Pei Huang; Zengguo Cao; Tianyi Zhang; Kaikai Jin; Meihui Liu; Yujie Bai; Zhiyuan Gong; Xuemeng Li; Yuanyuan Li; Haili Zhang; Hualei Wang

doi:10.1016/j.virs.2025.11.011

PDF
Cite
Share

facebook

twitter

google

linkedin

Volume 40 Issue 6

December 2025

Citation: Zanheng Huang, Pei Huang, Zengguo Cao, Tianyi Zhang, Kaikai Jin, Meihui Liu, Yujie Bai, Zhiyuan Gong, Xuemeng Li, Yuanyuan Li, Haili Zhang, Hualei Wang. Specific and visual detection of EBOV based on a one-pot RT-RAA-CRISPR/Cas12a assay .VIROLOGICA SINICA, 2025, 40(6) : 1054-1057. http://dx.doi.org/10.1016/j.virs.2025.11.011

Specific and visual detection of EBOV based on a one-pot RT-RAA-CRISPR/Cas12a assay

Zanheng Huang ^a ,
Pei Huang ^a ,
Zengguo Cao ^b ,
Tianyi Zhang ^a ,
Kaikai Jin ^a ,
Meihui Liu ^a ,
Yujie Bai ^a ,
Zhiyuan Gong ^a ,
Xuemeng Li ^c,d ,
Yuanyuan Li ^a ,
Haili Zhang ^{a
,,} ,
Hualei Wang ^{a
,,}

a. State Key Laboratory for Diagnosis and Treatment of Severe Zoonotic Infectious Diseases, Key Laboratory for Zoonosis Research of the Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine College of Veterinary Medicine, Jilin University, Changchun 130062, China;
b. Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Center for Biosafety Mega-Science, Chinese Academy of Sciences, Wuhan 430071, China;
c. Guangdong Provincial Key Laboratory of Medical Molecular Diagnostics, School of Basic Medicine, Guangdong Medical University, Dongguan 523000, China;
d. The Marine Biomedical Research Institute, Affiliated Hospital of Guangdong Medical University, Guangdong Medical University, Zhanjiang 524023, China

Corresponding author: Haili Zhang, zhanghaili@jlu.edu.cn
Hualei Wang, wanghualei@jlu.edu.cn
Received Date: 26 May 2025
Accepted Date: 26 November 2025
Available online: 28 November 2025

Abstract

Highlights
1. The one-pot RT-RAA-CRISPR/Cas12a assay for EBOV has single-copy resolution within 40 min.
2. This assay targeting the VP40 gene has high specificity for EBOV and no cross-reaction with SUDV.
3. This one-pot assay is easy to operate and can effectively avoid aerosol contamination.

Electronic Supplementary Material

10.1016j.virs.2025.11.011-ESM1.docx
References
1. Broadhurst, M.J., Brooks, T.J.G., Pollock, N.R., 2016. Diagnosis of Ebola virus disease: past, present, and future. Clin. Microbiol. Rev. 29, 773-793.
2. Bonney, L.C., Watson, R.J., Slack, G.S., Bosworth A, Wand, NIV., Hewson, R., 2020. A flexible format LAMP assay for rapid detection of Ebola virus. PLoS. Negl. Trop. Dis. 14(7):e0008496.
3. Chams, N., Chams, S., Badran, R., Shams, A., Araji, A., Raad, M., Mukhopadhyay, S., Stroberg, E., Duval, E.J., Barton, L.M., Hajj Hussein, I., 2020. COVID-19: a multidisciplinary review. Front. Public Health 8, 383.
4. Jacob, S.T., Crozier, I., Fischer, W.A., Hewlett, A., Kraft, C.S., Vega, M.-A. de L., Soka, M.J., Wahl, V., Griffiths, A., Bollinger, L., Kuhn, J.H., 2020. Ebola virus disease. Nat. Rev. Dis. Primers 6, 13.
5. Kaushik, A., Tiwari, S., Dev Jayant, R., Marty, A., Nair, M., 2016. Towards detection and diagnosis of Ebola virus disease at point-of-care. Biosens. Bioelectron. 75, 254-272.
6. Mukadi-Bamuleka, D., Mambu-Mbika, F., De Weggheleire, A., Edidi-Atani, F., Bulabula-Penge, J., Mfumu, M.M.K., Legand, A., Nkuba-Ndaye, A., N’kasar, Y.T.T., Mbala-Kingebeni, P., Klena, J.D., Montgomery, J.M., Muyembe-Tamfum, J.-J., Formenty, P., van Griensven, J., Arien, K.K., Ahuka-Mundeke, S., 2023. Efficiency of field laboratories for Ebola virus disease outbreak during chronic insecurity, Eastern democratic Republic of the Congo, 2018-2020. Emerg. Infect. Dis. 29, 1-9.
7. Mulangu, S., Dodd, L.E., Davey, R.T., Tshiani Mbaya, O., Proschan, M., Mukadi, D., Lusakibanza Manzo, M., Nzolo, D., Tshomba Oloma, A., Ibanda, A., Ali, R., Coulibaly, S., Levine, A.C., Grais, R., Diaz, J., Lane, H.C., Muyembe-Tamfum, J.-J., The Palm Writing Group, 2019. A randomized, controlled trial of Ebola virus disease therapeutics. N. Engl. J. Med. 381, 2293-2303.
Proportional views

Figures(1)

PDF

Article Metrics

Article views(1812) PDF downloads(17) Cited by()

Proportional views

Specific and visual detection of EBOV based on a one-pot RT-RAA-CRISPR/Cas12a assay

Corresponding author: Haili Zhang, zhanghaili@jlu.edu.cn

Corresponding author: Hualei Wang, wanghualei@jlu.edu.cn

a. State Key Laboratory for Diagnosis and Treatment of Severe Zoonotic Infectious Diseases, Key Laboratory for Zoonosis Research of the Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine College of Veterinary Medicine, Jilin University, Changchun 130062, China;
b. Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Center for Biosafety Mega-Science, Chinese Academy of Sciences, Wuhan 430071, China;
c. Guangdong Provincial Key Laboratory of Medical Molecular Diagnostics, School of Basic Medicine, Guangdong Medical University, Dongguan 523000, China;
d. The Marine Biomedical Research Institute, Affiliated Hospital of Guangdong Medical University, Guangdong Medical University, Zhanjiang 524023, China

Received Date: 26 May 2025
Accepted Date: 26 November 2025

Abstract: Highlights
1. The one-pot RT-RAA-CRISPR/Cas12a assay for EBOV has single-copy resolution within 40 min.
2. This assay targeting the VP40 gene has high specificity for EBOV and no cross-reaction with SUDV.
3. This one-pot assay is easy to operate and can effectively avoid aerosol contamination.