Performance evaluation of the automated SNPs analyzer for the detection of JAK2 V617F mutation

Abstract

Objective To test and evaluate the JAK2 gene V617F mutation in patients with myeloproliferative tumors based on i-densy IS-5320 platform according to ISO15189 accreditation requirements. Methods Instrument performance verification. Selected from December 2014 to February 2017, 20 cases of JAK2 V617F mutation positive peripheral blood samples from Huashan Hospital of the Shanghai FuDan University Medical College and 20 cases of peripheral blood samples with negative JAK2V617F mutation.The Realtime PCR with TaqMan MGB probe was selected as the control method to verify and evaluate the accuracy of testing JAK2 V617F mutation on i-densy IS-5320 platform. Whole blood samples were used to evaluate the reproducibility, cross-contamination and anti-interference ability of this platform. The ability of mutation load was verified by detecting mixtures of human erythnoleukemia cells and colorectal cancer cell HCT116 with 12 different proportions. Results I-densy IS-5320 platform and TaqMan MGB probe real-time fluorescence quantitative PCR show the same result.The within-run reproducibility and between-run reproducibility are both 100%. There is no observed contamination. High bilirubin and high triglyceride blood samples have no obvious interference on mutation detection. The mutation ratio with a load as low as 0.25% could be tested stably by i-densy IS-5320 platform. The detecting peak of melting curve can reflect the ratio of JAK2 V617F mutation to some extent. Conclusions I-densy IS-5320 can detect the mutation of JAK2 V617F gene in the whole blood directly. It has high sensitivity, accuracy and stability, and is easy to operate, and also can reflect the mutation load of JAK2 V617F, which could meet the clinical requirements for the detection of mutations. (Chin J Lab Med, 2018, 41: 470-474) Key words: Myeloproliferative neoplasms; JAK2 V617F; High resolution melting curve analysis