Delta-PCR as an efficient and sensitive method for measurement of MRD in AML patients with FLT3-ITD mutation

سال انتشار: 1397
نوع سند: مقاله کنفرانسی
زبان: انگلیسی
مشاهده: 425

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شناسه ملی سند علمی:

IPMCMED03_015

تاریخ نمایه سازی: 6 خرداد 1398

چکیده مقاله:

Introduction FLT3 internal tandem duplications (FLT3/ITDs) are reported in acute myeloid leukemia(AML) and predict poor clinical outcome. This mutation is present in about 20-30% of patients with AML. The use of methods with a higher sensitivity and specificity than conventional methods for detecting minor clones at the time of initial diagnosis and for measuring the minimal residual disease (MRD) in mutated patients is important. Delta PCR is a new method that can increase the specificity of PCR by using three primers simultaneously. MethodBone marrow and/or blood samples were collected from 31 patients with AML, whose FLT3-ITD mutation was confirmed standard fragment analysis, At the diagnosis, and then at intervals of 1-3 months. Mononuclear cells were separated by centrifugation through Ficoll-Hypaque and The DNAs were extracted by the salting out method. Delta PCR method was used for detection of MRD during follow up times. Two pairs of PCR product with the 19 + 1 Delta were interpreted as ITD-positive. After data collection, t-test and x2 were used to analyze the data. To determine the sensitivity of the method, a DNA sample with FLT3-ITD allele burden 50% was diluted in different concentration in a normal DNA sampleResultsThe Delta PCR method confirmed the presence of mutations in all studies samples. The Delta PCR method was able to detect MRD 25 days ( range 20 - 45 days) before recurrence of disease . The sensitivity of the method was 1 in 200 (0.5%). ConclusionThe Delta PCR method was able to detect all ITD mutations in patients. In addition, due to the simultaneous multiplication of two bands, the specificity of this method is more than the standard fragment analysis method, which is done using a pair of primers. In addition, by optimizing the conditions of electrophoresis, the sensitivity of this method can be increased to some extent.

نویسندگان

Maryam Arab

Allied medical sciences school, Tehran University of Medical Sciences, Tehran, Iran

Bahram Chahardouli

Hematology-Oncology & Stem cell transplantation Research center, Tehran University of Medical Sciences, Tehran, Iran

Shahrbano Rostami

Hematology-Oncology & Stem cell transplantation Research center, Tehran University of Medical Sciences, Tehran, Iran

Arvin Samemaleki

Allied medical sciences school, Tehran University of Medical Sciences, Tehran, Iran