A clustered regularly interspaced palindromic repeat-Cas12a-based assay for rapid dengue virus monitoring in mosquito and clinical samples facilitating global collaborative surveillance and risk assessment to block dengue spreading

Xiang Guo; Minling Hu; Ying Lu; Xiaohua Liu; Haiyang Chen; Xiaoqing Zhang; Liu Ge; Ziyao Li; Yuji Wang; Qing He; Shu Zeng; Wenwen Ren; Lingzhai Zhao; Zhiqiang Peng; Wushen Chen; Chunmei Wang; Qingming Kong; Shaohong Lu; Fuchun Zhang; Lei Luo; Hua Li; Xiao-Guang Chen; Xiaohong Zhou

doi:10.1002/inmd.70074

Interdisciplinary Medicine ›› 2026, Vol. 4 ›› Issue (1) :e70074 DOI: 10.1002/inmd.70074

RESEARCH ARTICLE

A clustered regularly interspaced palindromic repeat-Cas12a-based assay for rapid dengue virus monitoring in mosquito and clinical samples facilitating global collaborative surveillance and risk assessment to block dengue spreading

Author information +

¹. Department of Pathogen Biology, Institute of Tropical Medicine, School of Public Health, Guangdong Provincial Key Laboratory of Tropical Disease Research, Key Laboratory of Prevention and Control for Emerging Infectious Diseases of Guangdong Higher Institutes, Key Laboratory of Infectious Diseases Research in South China of Ministry of Education, Southern Medical University, Guangzhou, China

². School of Basic Medical Sciences, Henan University, Kaifeng, China

³. Guangzhou Center for Disease Control and Prevention, Guangzhou, China

⁴. School of Basic Medicine and Forensics, Key Laboratory of Bio-tech Vaccine of Zhejiang Province, Engineering Research Center of Novel Vaccine of Zhejiang Province, Zhejiang Provincial People's Hospital (Affiliated People's Hospital), Hangzhou Medical College, Hangzhou, China

⁵. Guangzhou Medical Research Institute of Infectious Diseases, Infectious Disease Center, Guangzhou Eighth People's Hospital, Guangzhou Medical University, Guangzhou, China

⁶. Guangdong Provincial Center for Disease Control and Prevention, Guangzhou, China

⁷. Shenzhen Luohu Center for Disease Control and Prevention, Shenzhen, China

⁸. Experimental Teaching Center of Preventive Medicine, National Experimental Teaching Demonstration Center of Preventive Medicine, Southern Medical University, Guangzhou, China

llyeyq@163.com

smumph@163.com

daizhouxh@163.com

Show less

History +

PDF

Abstract

Dengue fever, an acute Aedes-borne infectious disease caused by the dengue virus (DENV), is a substantial global health challenge, necessitating the development of swift and effective surveillance methodologies. We have developed a Recombinase Aided Amplification (RAA)–Cas12a assay that exhibits superior analytical sensitivity compared with real-time PCR while maintaining comparable specificity. In clinical specimen testing, the assay demonstrated a sensitivity of 92.3% and a specificity of 100%, using a signal-to-noise ratio of ≥2.0 as the positivity threshold. Furthermore, the RAA–Cas12a assay displayed remarkable sensitivity in detecting DENV in both adult and larval mosquitoes, with field data from mosquito surveillance aligning closely with the emergence of local dengue cases. Characterized by its simplicity, rapidity, and precision, the RAA–Cas12a approach emerges as a highly promising technology for monitoring virus carriage in field-collected mosquitoes, thereby functioning as both an early warning system and a crucial indicator for effective dengue outbreak management. The practicality and cost-effectiveness of this methodology provide a robust foundation for the establishment of an efficient global collaborative surveillance and risk assessment platform to mitigate the rapid spread of dengue.

Keywords

Aedes albopictus / collaborative surveillance / CRISPR/Cas12a / dengue virus (DENV) / risk assessment

Cite this article

Download citation ▾

Xiang Guo, Minling Hu, Ying Lu, Xiaohua Liu, Haiyang Chen, Xiaoqing Zhang, Liu Ge, Ziyao Li, Yuji Wang, Qing He, Shu Zeng, Wenwen Ren, Lingzhai Zhao, Zhiqiang Peng, Wushen Chen, Chunmei Wang, Qingming Kong, Shaohong Lu, Fuchun Zhang, Lei Luo, Hua Li, Xiao-Guang Chen, Xiaohong Zhou. A clustered regularly interspaced palindromic repeat-Cas12a-based assay for rapid dengue virus monitoring in mosquito and clinical samples facilitating global collaborative surveillance and risk assessment to block dengue spreading. Interdisciplinary Medicine, 2026, 4(1): e70074 DOI:10.1002/inmd.70074

登录浏览全文

4963

注册一个新账户忘记密码

References

Publishing order | Descend order by publishing year | Descend order by cited within

[1]	M. G. Guzman, D. J. Gubler, A. Izquierdo, E. Martinez, S. B. Halstead, Nat. Rev. Dis. Primers 2016, 2, 16055.

[2]

M. U. G. Kraemer, R. C. Reiner., O. J. Brady, J. P. Messina, M. Gilbert, D. M. Pigott, D. Yi, K. Johnson, L. Earl, L. B. Marczak, S. Shirude, N. Davis Weaver, D. Bisanzio, T. A. Perkins, S. Lai, X. Lu, P. Jones, G. E. Coelho, R. G. Carvalho, W. Van Bortel, C. Marsboom, G. Hendrickx, F. Schaffner, C. G. Moore, H. H. Nax, L. Bengtsson, E. Wetter, A. J. Tatem, J. S. Brownstein, D. L. Smith, L. Lambrechts, S. Cauchemez, C. Linard, N. R. Faria, O. G. Pybus, T. W. Scott, Q. Liu, H. Yu, G. R. W. Wint, S. I. Hay, N. Golding, Nat. Microbiol. 2019, 4, 854.

[3]	S. Bhatt, P. W. Gething, O. J. Brady, J. P. Messina, A. W. Farlow, C. L. Moyes, J. M. Drake, J. S. Brownstein, A. G. Hoen, O. Sankoh, M. F. Myers, D. B. George, T. Jaenisch, G. R. Wint, C. P. Simmons, T. W. Scott, J. J. Farrar, S. I. Hay, Nature 2013, 496, 504.

[4]	World Health Organization, WHO launches global strategic plan to fight rising dengue and other Aedes-borne arboviral diseases, https://www.who.int/news/item/03-10-2024-who-launches-global-strategic-plan-to-fight-rising-dengue-and-other-aedes-borne-arboviral-diseases (accessed October 2024).

[5]	World Health Organization, Dengue: guidelines for diagnosis, treatment, prevention and control. WHO Press – World Health Organization, Geneve, Switzerland2009.

[6]	Q. A. Ten Bosch, H. E. Clapham, L. Lambrechts, V. Duong, P. Buchy, B. M. Althouse, A. L. Lloyd, L. A. Waller, A. C. Morrison, U. Kitron, G. M. Vazquez-Prokopec, T. W. Scott, T. A. Perkins, PLoS Pathog. 2018, 14, e1006965.

[7]	R. W. Peeling, H. Artsob, J. L. Pelegrino, P. Buchy, M. J. Cardosa, S. Devi, D. A. Enria, J. Farrar, D. J. Gubler, M. G. Guzman, S. B. Halstead, E. Hunsperger, S. Kliks, H. S. Margolis, C. M. Nathanson, V. C. Nguyen, N. Rizzo, S. Vazquez, S. Yoksan, Nat. Rev. Microbiol. 2010, 8, S30.

[8]	L. Li, X. Guo, X. Zhang, L. Zhao, L. Li, Y. Wang, T. Xie, Q. Yin, Q. Jing, T. Hu, Z. Li, R. Wu, W. Zhao, S. X. Xin, B. Shi, J. Liu, S. Xia, Z. Peng, Z. Yang, F. Zhang, X. G. Chen, X. Zhou, Infect. Dis. Poverty 2022, 11, 107.

[9]

L. Zhao, X. Guo, L. Li, Q. Jing, J. Ma, T. Xie, D. Lin, L. Li, Q. Yin, Y. Wang, X. Zhang, Z. Li, X. Liu, T. Hu, M. Hu, W. Ren, J. Li, J. Peng, L. Yu, Z. Peng, W. Hong, X. Leng, L. Luo, J. J. K. Ngobeh, X. Tang, R. Wu, W. Zhao, B. Shi, J. Liu, Z. Yang, X. G. Chen, X. Zhou, F. Zhang, Infect. Dis. Poverty 2024, 13, 43.

[10]	X. Guo, P. Teng, Z. Li, L. Li, X. Liu, X. Zhang, Y. Wang, M. Hu, W. Ren, S. Zeng, H. Chen, L. Ge, S. Liu, Z. Peng, J. Sun, X. Zhang, L. Luo, J. Peng, B. Shi, R. Wu, J. Liu, F. Zhang, X.-G. Chen, T. Chen, X. Zhou, iMetaOmics 2025, 2, e70048.

[11]	M. M. Kaminski, O. O. Abudayyeh, J. S. Gootenberg, F. Zhang, J. J. Collins, Nat. Biomed. Eng. 2021, 5, 643.

[12]

C. Myhrvold, C. A. Freije, J. S. Gootenberg, O. O. Abudayyeh, H. C. Metsky, A. F. Durbin, M. J. Kellner, A. L. Tan, L. M. Paul, L. A. Parham, K. F. Garcia, K. G. Barnes, B. Chak, A. Mondini, M. L. Nogueira, S. Isern, S. F. Michael, I. Lorenzana, N. L. Yozwiak, B. L. MacInnis, I. Bosch, L. Gehrke, F. Zhang, P. C. Sabeti, Science 2018, 360, 444.

[13]	J. S. Gootenberg, O. O. Abudayyeh, J. W. Lee, P. Essletzbichler, A. J. Dy, J. Joung, V. Verdine, N. Donghia, N. M. Daringer, C. A. Freije, C. Myhrvold, R. P. Bhattacharyya, J. Livny, A. Regev, E. V. Koonin, D. T. Hung, P. C. Sabeti, J. J. Collins, F. Zhang, Science 2017, 356, 438.

[14]	J. P. Broughton, X. Deng, G. Yu, C. L. Fasching, V. Servellita, J. Singh, X. Miao, J. A. Streithorst, A. Granados, A. Sotomayor-Gonzalez, K. Zorn, A. Gopez, E. Hsu, W. Gu, S. Miller, C. Y. Pan, H. Guevara, D. A. Wadford, J. S. Chen, C. Y. Chiu, Nat. Biotechnol. 2020, 38, 870.

[15]	J. S. Chen, E. Ma, L. B. Harrington, M. Da Costa, X. Tian, J. M. Palefsky, J. A. Doudna, Science 2018, 360, 436.

[16]	L. Li, S. Li, N. Wu, J. Wu, G. Wang, G. Zhao, J. Wang, ACS Synth. Biol. 2019, 8, 2228.

[17]	S. Y. Li, Q. X. Cheng, J. M. Wang, X. Y. Li, Z. L. Zhang, S. Gao, R. B. Cao, G. P. Zhao, J. Wang, Cell Discov. 2018, 4, 20.

[18]	C. M. Ackerman, C. Myhrvold, S. G. Thakku, C. A. Freije, H. C. Metsky, D. K. Yang, S. H. Ye, C. K. Boehm, T. F. Kosoko-Thoroddsen, J. Kehe, T. G. Nguyen, A. Carter, A. Kulesa, J. R. Barnes, V. G. Dugan, D. T. Hung, P. C. Blainey, P. C. Sabeti, Nature 2020, 582, 277.

[19]	R. A. Lee, H. Puig, P. Q. Nguyen, N. M. Angenent-Mari, N. M. Donghia, J. P. McGee, J. D. Dvorin, C. M. Klapperich, N. R. Pollock, J. J. Collins, Proc. Natl. Acad. Sci. USA 2020, 117, 25722.

[20]	W. Chen, H. Luo, L. Zeng, Y. Pan, J. B. Parr, Y. Jiang, C. H. Cunningham, K. L. Hawley, J. D. Radolf, W. Ke, J. Ou, J. Yang, B. Yang, H. Zheng, Nat. Commun. 2022, 13, 4671.

[21]	L. A. Curti, F. Pereyra-Bonnet, G. D. Repizo, J. V. Fay, K. Salvatierra, M. J. Blariza, D. Ibanez-Alegre, A. R. Rinflerch, M. Miretti, C. A. Gimenez, Emerg. Microbes Infect. 2020, 9, 1140.

[22]	J. S. Gootenberg, O. O. Abudayyeh, M. J. Kellner, J. Joung, J. J. Collins, F. Zhang, Science 2018, 360, 439.

[23]	M. Zhong, J. Liu, J. Wu, J. Li, N. Luo, C. Zhu, R. Liu, Q. Xia, H. Ju, Sensor. Actuator. B Chem. 2022, 366, 132011.

[24]	G. Tian, J. Tan, B. Liu, M. Xiao, Q. Xia, Anal. Chim. Acta 2024, 1316, 342838.

[25]	T. Ayllon, R. M. Campos, P. Brasil, F. C. Morone, D. C. P. Camara, G. L. S. Meira, E. Tannich, K. A. Yamamoto, M. S. Carvalho, R. S. Pedro, J. Schmidt-Chanasit, D. Cadar, D. F. Ferreira, N. A. Honorio, Emerg. Infect. Dis. 2017, 23, 1411.

[26]	K. Kesorn, P. Ongruk, J. Chompoosri, A. Phumee, U. Thavara, A. Tawatsin, P. Siriyasatien, PLoS One 2015, 10, e0125049.

[27]	M. R. Donalisio, A. R. R. Freitas, A. Zuben, Rev. Saude Publica 2017, 51, 30.

[28]

M. Patchsung, K. Jantarug, A. Pattama, K. Aphicho, S. Suraritdechachai, P. Meesawat, K. Sappakhaw, N. Leelahakorn, T. Ruenkam, T. Wongsatit, N. Athipanyasilp, B. Eiamthong, B. Lakkanasirorat, T. Phoodokmai, N. Niljianskul, D. Pakotiprapha, S. Chanarat, A. Homchan, R. Tinikul, P. Kamutira, K. Phiwkaow, S. Soithongcharoen, C. Kantiwiriyawanitch, V. Pongsupasa, D. Trisrivirat, J. Jaroensuk, T. Wongnate, S. Maenpuen, P. Chaiyen, S. Kamnerdnakta, J. Swangsri, S. Chuthapisith, Y. Sirivatanauksorn, C. Chaimayo, R. Sutthent, W. Kantakamalakul, J. Joung, A. Ladha, X. Jin, J. S. Gootenberg, O. O. Abudayyeh, F. Zhang, N. Horthongkham, C. Uttamapinant, Nat. Biomed. Eng. 2020, 4, 1140.

[29]	X. Wang, Q. Rao, Z. Lu, X. Deng, R. Shen, R. Wang, W. Dong, X. Qi, Z. Jin, Y. Tang, D. Du, Sci. Bull. 2023, 68, 3142.

[30]	M. A. J. Golding, S. A. A. Noble, N. K. Khouri, R. N. A. Layne-Yarde, I. Ali, S. L. Sandiford, Parasit Vectors 2023, 16, 442.

[31]	S. Lequime, R. E. Paul, L. Lambrechts, PLoS Pathog. 2016, 12, e1005548.

[32]	D. Xia, X. Guo, T. Hu, L. Li, P. Y. Teng, Q. Q. Yin, L. Luo, T. Xie, Y. H. Wei, Q. Yang, S. K. Li, Y. J. Wang, Y. Xie, Y. J. Li, C. M. Wang, Z. C. Yang, X. G. Chen, X. H. Zhou, Infect. Dis. Poverty 2018, 7, 89.

[33]	J. Peccia, A. Zulli, D. E. Brackney, N. D. Grubaugh, E. H. Kaplan, A. Casanovas-Massana, A. I. Ko, A. A. Malik, D. Wang, M. Wang, J. L. Warren, D. M. Weinberger, W. Arnold, S. B. Omer, Nat. Biotechnol. 2020, 38, 1164.

[34]

F. Amman, R. Markt, L. Endler, S. Hupfauf, B. Agerer, A. Schedl, L. Richter, M. Zechmeister, M. Bicher, G. Heiler, P. Triska, M. Thornton, T. Penz, M. Senekowitsch, J. Laine, Z. Keszei, P. Klimek, F. Nagele, M. Mayr, B. Daleiden, M. Steinlechner, H. Niederstatter, P. Heidinger, W. Rauch, C. Scheffknecht, G. Vogl, G. Weichlinger, A. O. Wagner, K. Slipko, A. Masseron, E. Radu, F. Allerberger, N. Popper, C. Bock, D. Schmid, H. Oberacher, N. Kreuzinger, H. Insam, A. Bergthaler, Nat. Biotechnol. 2022, 40, 1814.