Tuyển tập báo cáo các nghiên cứu khoa học quốc tế ngành y học dành cho các bạn tham khảo đề tài: Development of a Highly Sensitive Method for Detection of JAK2V617F. | Zhao et al. Journal of Hematology Oncology 2011 4 40 http content 4 1 40 JOURNAL OF HEMATOLOGY ONCOLOGY SHORT REPORT Open Access Development of a Highly Sensitive Method for Detection of JAK2V617F Anna H Zhao1 2 Rufei Gao1 3 and Zhizhuang J Zhao1 3 Abstract Background Ph- myeloproliferative neoplasms MPNs represent a heterogeneous group of chronic diseases characterized by increased expansion of hematopoietic cells of the myeloid lineage. JAK2V617F an activation mutation form of tyrosine kinase JAK2 is found in the majority of patients with MPNs. Studies have demonstrated that JAK2V617F can cause MPNs and various methods have been developed to detect JAK2V617F for diagnostic purposes. However a highly sensitive method is still needed for the earliest possible detection and for disease prevention and treatment. Methods In the present study we developed a method dubbed restriction fragment nested allele-specific PCR RFN-AS-PCR . The method consists of three steps 1 initial amplification of DNA samples with PCR primers surrounding the JAK2V617F mutation site 2 digestion of the PCR products with restriction enzyme BsaXI which only cleaves the wild type allele and 3 detection of JAK2V617F by allele-specific PCR with nested primers. Results We tested the sensitivity of the method by using purified plasmid DNAs and blood cell DNAs containing known proportions of JAK2V617F. We were able to detect JAK2V617F with a sensitivity of . We further analyzed blood cell DNA samples from 105 healthy donors with normal blood cell counts and found three JAK2V617F-positive cases which would have remained undetected using a less sensitive method. Conclusions We have developed a highly sensitive method that will allow for detection of JAK2V617F at a very early stage. This method may have major implications in diagnosis and prevention of MPNs and related diseases. Keywords Tyrosine kinase myeloproliferative neoplasms JAK2 mutation detection diagnosis Background Ph-
