[Detection of JAK2V617F and CALR Gene Mutations by Multiplex of Patients with Myeloproliferative Neoplasms PCR-Capillary Electrophoresis].
作者信息
Yuan Jian-Long, Shi Ying-Xu, DU Hua, Wang Ying-Jun, Zhao Zi-Ling, Li Ge, Han Yan-Qiu
机构信息
Department of Clinical Laboratory Examination, Affiliated Hospital of Inner Mongolia Medical University,Huhhot 010050, Inner Mongolia Autonomous Region, China.
Department of Pathology, Affiliated Hospital of Inner Mongolia Medical University,Huhhot 010050, Inner Mongolia Autonomous Region, China,Basic Medical College, Inner Mongolia Medical University,Huhhot 010050, Inner Mongolia Autonomous Region, China.
出版信息
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2020 Dec;28(6):1998-2003. doi: 10.19746/j.cnki.issn.1009-2137.2020.06.033.
OBJECTIVE
To evaluate the proformance of multiplex PCR and capillary electrophoresis(MPCE) in the detection of JAK2V617F and CALR mutation in myeloproliferative neoplasms(MPN).
METHODS
The specificity primers of JAK2617F gene mutation and the primers of CALR gene were designed at the same time. The JAK2V617F and CALR gene primers were labeled with Cy5 fluorescence, all the primers were mixed in one tube for multiplex PCR and the PCR prodcuts were analysised by capillary electrophoresis. Then detection limit and sensitivity of MPCE were evaluated, and compared with comercial diagnostic kit.
RESULTS
JAK2V617F and CALR gene mutations could be detect by MPCE in one PCR test. JAK2V617F mutation could be detected at 0.01 ng genomic DNA, double positive JAK2V617F and CLAR gene mutations could be detected at 0.1 ng genomic DNA, at least 0.1% JAK2V617F positive mutation could be detected. The consistency between MPCE and commercial diagnostic gene mutation kit was 100%.
CONCLUSION
It is developed that a new gene mutation detection method of JAK2 V617F and CLAR gene based on MPCE in our experiment and it can be used as a new reagent for molecular diagnosis of MPN patients.
Zotero 插件