Diagnotic procedure development to detect common point mutations in fms-like tyrosine kinase 3 tyrosine kinase domain (flt3-tkd)

Do Thi Thanh Trung, Nguyen Mai Phuong, Trinh Le Phuong, Pham Bao Yen

Abstract


Acute myeloid leukemia (AML) is one of the high mortality-rate malignant hematological diseases. Mutations of tyrosine kinase receptors, including FMS-like tyrosine kinase 3 (FLT3), are involved in the constitutive enzymatic activation and abnormal differentiation of leukocytes as characterized in AML patients. The two most common FLT3 mutations, internal tandem duplications (ITD) and tyrosine kinase domain (TKD) single nucleotide substitutions, are considered poor prognosis factors affecting treatment decision for AML patients. Duplications of the juxtamembrane domain are found in 20-25% of AML patients with normal karyotype, while point mutations in the activation loop located within TKD are diagnosed in 7% cases. This study has successfully established and optimized a procedure to detect FLT3-TKD point mutations in codon 835 and 836 coding aspartic acid and isoleucine (D835 and I836, respectively) in 4 steps: (1) Amplification of a 114 bp - fragment of FLT3 gene with specific primers using polymerase chain reaction (PCR); (2) Digestion of the amplified fragment by EcoRV. (3) Electrophoresis of the digested products on 12% polyacrylamide gel; (4) Comparison of band pattern with the mutant and normal controls both generated in our laboratory. Preliminary screening of 126 Vietnamese AML patients revealed five mutations (4.0%), replacing G for T in codon 835.

Keywords


Acute myeloid leukemia (AML), D835 mutations, FMS-like tyrosine kinase 3 (FLT3), FLT3-TKD, I835 mutations

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DOI: https://doi.org/10.15625/0866-7160/v37n1se.6088 Display counter: Abstract : 51 views. Untitled : 18 views.

 

                 

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