Dihydrorhodamine-123 flow cytometry method: time for substantial revision in technical procedure.

The dihydrorhodamine 123 assay is generally applied to measure the production of intracellular reactive oxygen species in neutrophils using flow cytometry and is considered a diagnostic evaluation for chronic granulomatous disease. In fact, there is a broad range of variables that can directly or indirectly affect test results, either individually or collectively. It is therefore crucial to identify the ideal requirements to achieve reliable results as well as using these requirements to provide standard operating procedures that should be taken into account. Therefore, we focus on aligning optimum results by comparing preanalytical and analytical phases that influence test results, such as the effect of various anticoagulants, transport and maintaining temperature (24°C or 4°C) of samples, test prime run time, appropriate solution concentrations, and effect of incubation temperature (24°C or 37°C) during the test run.

Mixed-phenotype acute leukemia characteristics: first report from Iran.

Mixed-phenotype acute leukemia (MPAL) is the infrequent type of acute leukemia characterized by immunophenotypic and/or cytochemical features of both lineages, but the diagnosis of this disease still is a challenge. In this study, we analyzed immunophenotyping, cytochemistry and frequency of MPAL patients to better diagnosis of MPAL characteristics according to WHO 2016 criteria for the first time in Iran. In this retrospective study, 27 patients were diagnosed as MPAL based on WHO 2016 criteria during 2014-2017. Flow cytometric immunophenotyping was performed on PB and BM samples evaluation of different CD marker expressions in MPAL subsets. RT-PCR was performed for the analyses of BCR/ABL1 fusion in MPAL subsets. Among 27 cases, (70.4%) 19 cases were B + My, (22.22%) 6 cases were T + My, and 2 cases (7.40%) were B + T + My. CD34, CD19, HLA-DR, TdT, CD22, iMPO were positive in majority of B + My cases. CD45, iMPO, iCD3, CD7, CD2 and CD5 were positive in majority of T + My cases. HLA-DR, TdT, CD10, CD22, iCD79a, iMPO, CD45, iCD3, CD7, CD3, CD2, CD5 were positive in majority of B + T + My cases. BCR/ABL1 fusion was positive for 3 cases (11.1%) of p190 fusion and 2 cases (7.4%) of p210 fusion in B + My cases. WHO 2016 criteria are the current standard for diagnosing MPAL. Also, evaluation of TdT, CD2, CD5, CD7 expressions by flow cytometry in EGIL criteria is useful for the better diagnosis of MPAL subsets. In addition, evaluation of BCR/ABL1 and MLL rearrangements in patients should be part of standard work-up in MPAL.