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Cas12a and Lateral Flow Strip-Based Test for Rapid and Ultrasensitive Detection of Spinal Muscular Atrophy
Article

CRISPR/Cas12a-Based Ultrasensitive and Rapid Detection of JAK2 V617F Somatic Mutation in Myeloproliferative Neoplasms

Center for Medical Genetics & Hunan Key Laboratory of Medical Genetics, School of Life Sciences, Central South University, Changsha 410078, China
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Authors to whom correspondence should be addressed.
Received: 2 July 2021 / Revised: 20 July 2021 / Accepted: 22 July 2021 / Published: 24 July 2021
(This article belongs to the Special Issue Application of CRISPR Cas Systems for Biosensing)
The JAK2 V617F mutation is a major diagnostic, therapeutic, and monitoring molecular target of Philadelphia-negative myeloproliferative neoplasms (MPNs). To date, numerous methods of detecting the JAK2 V617F mutation have been reported, but there is no gold-standard diagnostic method for clinical applications. Here, we developed and validated an efficient Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR associated protein 12a (Cas12a)-based assay to detect the JAK2 V617F mutation. Our results showed that the sensitivity of the JAK2 V617F/Cas12a fluorescence detection system was as high as 0.01%, and the JAK2 V617F/Cas12a lateral flow strip assay could unambiguously detect as low as 0.5% of the JAK2 V617F mutation, which was much higher than the sensitivity required for clinical application. The minimum detectable concentration of genomic DNA achieved was 0.01 ng/μL (~5 aM, ~3 copies/μL). In addition, the whole process only took about 1.5 h, and the cost of an individual test was much lower than that of the current assays. Thus, our methods can be applied to detect the JAK2 V617F mutation, and they are highly sensitive, rapid, cost-effective, and convenient. View Full-Text
Keywords: JAK2 V617F; CRISPR/Cas12a; mutation detection; fluorescence detection system; lateral flow strip; Philadelphia-negative myeloproliferative neoplasms JAK2 V617F; CRISPR/Cas12a; mutation detection; fluorescence detection system; lateral flow strip; Philadelphia-negative myeloproliferative neoplasms
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MDPI and ACS Style

Chen, M.; Zhang, C.; Hu, Z.; Li, Z.; Li, M.; Wu, L.; Zhou, M.; Liang, D. CRISPR/Cas12a-Based Ultrasensitive and Rapid Detection of JAK2 V617F Somatic Mutation in Myeloproliferative Neoplasms. Biosensors 2021, 11, 247. https://0-doi-org.brum.beds.ac.uk/10.3390/bios11080247

AMA Style

Chen M, Zhang C, Hu Z, Li Z, Li M, Wu L, Zhou M, Liang D. CRISPR/Cas12a-Based Ultrasensitive and Rapid Detection of JAK2 V617F Somatic Mutation in Myeloproliferative Neoplasms. Biosensors. 2021; 11(8):247. https://0-doi-org.brum.beds.ac.uk/10.3390/bios11080247

Chicago/Turabian Style

Chen, Miaomiao, Chunhua Zhang, Zhiqing Hu, Zhuo Li, Menglin Li, Lingqian Wu, Miaojin Zhou, and Desheng Liang. 2021. "CRISPR/Cas12a-Based Ultrasensitive and Rapid Detection of JAK2 V617F Somatic Mutation in Myeloproliferative Neoplasms" Biosensors 11, no. 8: 247. https://0-doi-org.brum.beds.ac.uk/10.3390/bios11080247

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