PCR đặc hiệu allele tích hợp công nghệ amplification refractory mutation system (Dsa-Pcr-Arms) phát hiện đột biến jak2 v617f và Carl trên bệnh nhân có hội chứng tăng sản tủy

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BIDIRECTIONAL ALLELE SPECIFIC PCR INTEGRATED WITH AMPLIFICATION REFRACTORY MUTATION SYSTEM –(DSA-PCR-ARMS) FOR ANALYZING CARL AND JAK2 V617F MUTATIONS IN PATIENTS WITH MYELOPROLIFERATIVE NEOPLASM Bachground: Jak2 V617F and CARLmutation arerecognized as essential molecular genetic markers in diagnostic definition of myeloid proliferative neoplasm (MPN). Objective: to establish an income relevant Allele-specific-PCR-ARMS (DSA-PCR-ARMS) assays to identify Jak2 V617F, CARL mutations in MPN patients. Subjects: 64 MPN patients’ samples, one V617F positive referrance samples and one CALR mutant positive sample. Methods: dilution series of 100%, 50%, 25%, 5%, 0,5% and 0% mutant alleles were created as pseudo-samples establish the detection limits and the technical specificity of DSA-PCR-ARMS assays in identifying the mentioned genetics markers. Result: Our approach can detect presence of 0.5 % V617F mutant DNA fragment; beside that just by one gel-based PCR reaction, we can differentiate the deletion/insertion CARL carriers from the wild type counterpart. By applying our approachesto a study cohort of 50 MPN cases, a sensitivity of 64% and specificity of 100% was recorded for our Jak2 V617F detection assay and we also detected 3 CARL mutation case amongst the 10 V617F negative cases. Conclusion: The in-house of Jak2 V617F/CARL mutation detection protocols have been successfully established in our hospital. Key words: Jak2 V617F, CARL, myeloid proliferative neoplasm (MPN).