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Highly Sensitive Droplet Digital PCR for Detection of Low-level JAK2V617F Mutation |
LONG Ling-yu1,ZHOU Ya-lan1,**(),JIAO Yao1,YE Lin-hai2,LI Xiu-chun2,LIU Yan-rong1,HUANG Xiao-jun1,3,RUAN Guo-rui1,**() |
1 Peking University People’s Hospital, Peking University Institute of Hematology, National Clinical Research Center for Hematologic Disease, Beijing Key Laboratory of Hematopoietic Stem Cell Transplantation, Peking University, Beijing 100044,China 2 Sniper Medical Technology Co., Ltd., Suzhou 215000,China 3 Peking-Tsinghua Center for Life Sciences, Academy for Advanced Interdisciplinary Studies, Peking University, Beijing 100084,China |
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Abstract Objective: To establish a droplet digital PCR (ddPCR) method for detecting the low-level JAK2V617F mutation and to explore its application value in the diagnosis of myeloproliferative neoplasm (MPN). Methods: Site-specific TaqMan-MGB probes were used to establish the ddPCR method for detecting the JAK2V617F mutation in genomic DNA. JAK2V617F mutations were detected by ddPCR, real-time quantitative PCR and next-generation sequencing (NGS). The consistency of the detection results of different methods was compared. Results: The ddPCR method for detecting the JAK2V617F mutation based on specific TaqMan-MGB probes was successfully established, and the detection sensitivity was at least 0.05%. The ddPCR method and qPCR method were used to detect the JAK2V617F mutation in 82 subjects with MPN and 8 healthy controls. The consistency rate was 96.7%. For the 52 samples with positive results by both methods, the correlation coefficient of quantitative data was 0.971 4 (P<0.000 1). Eleven samples were verified by NGS and the correlation coefficient between the two quantitative results was 0.983 9 (P<0.000 1). Conclusions: The ddPCR with the specific TaqMan-MGB probe method can detect the JAK2V617F mutation conveniently and accurately, which may have potential application value in the screening and dynamic monitoring of the JAK2V617F mutation.
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Received: 10 October 2023
Published: 16 January 2024
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