JAK2V617F Mutation Testing In Myeloproliferative Neoplasms

[Obsjective]:Myeloproliferative neoplasms (MPNs) originated from pluripotent hematopoietic stem cells. Its major feature is abnormal clonal proliferation of the marrow cells and the differentiation of one or more lineages of them is relatively mature.The morpha and function of the proliferating cells are relatively normal.It always shows that one or more series of cells(involving erythrocytic series,thrombocytic series and Granulocyte-monocyte cell series) which contained pluripotent myeloid hematopoietic stem cells hyperplasy malignantly.According to the proliferation of cellular components,this kind of diseases can be divided into the following four kinds:Polycythemia vera(PV),Essential thrombocythemia(ET),Chronic myeloid leukemia(CML)and primary myelofibrosis(PMF),which can be transformed into each other though their performances are different.A number of studies have recently shown that the12th exon in the JAK2gene of the patients with negative BCR-ABL exists a high frequency point mutations---JAK2V617F.It shows that the first base of the617th codon(GTC)exists G�'Tpoint mutation,which help the coded screenshot acid(GTC) changing into the phenylalanine acid(TTC).The gene mutation can be found in a lot of diseases,which can offer diagnostic basis to further clarify the broad spectrum of blood disease initiated by it.The article preliminary find out the distribution of JAK2V617F by detecting45samples,which offers basis for the clinical diagnosis and treatment.[Methods]:1.Extraction of genomic DNA:extract mononuclear cell genomic DNA from the peripheral blood (bone marrow) specimens.2.Amplified though polymerase chain reaction:designed and synthetized the amplified JAK2V617F gene specific primers, established and optimized the amplification system and amplification conditions of PCR, then amplified;3.Agarose gel electrophoresis identification:amplification products by agarose gel electrophoresis for identification, only one negative, two stripes appear as positive;4.Gene sequencing:positive samples after PCR amplification purification for genetic sequencing;[Results]:1.A total of12patients exist JAK2V617F mutation,7cases of PV (87.5%),4cases of ET (33.3%), leases of PMF (1/2);2. Gene sequencing results for the positive rate of PV group7/8(87.5%%),ET group4/12(33.3%), the PMF group1/23.It suggest that the two test methods of AML, ALL, CML, CNL, CEL patients with hematologic malignancies and normal controls were negative.[Conclusion]:1.BCR-ABL negative patients of MPNs had JAK2V617F point mutation,the rest hadn’t, such as AML、ALL、CNLand CML。2.There are significant differences about the distribution of JAK2V617F in patients with PV, ET、PMF(P<0.05);3.JAK2V617F can be characteristic events of molecular for PV, ET、PMF and can be an independent indicator for clinical diagnosis of PV、ET、PMF;4、AS-PCR worth carrying out clinical testing methods。...