38 research outputs found
Laboratory recommendations for scoring deep molecular responses following treatment for chronic myeloid leukemia
No full textTreatment of chronic myeloid leukemia (CML) with tyrosine kinase inhibitors has advanced to a stage where many patients achieve very low or undetectable levels of disease. Remarkably, some of these patients remain in sustained remission when treatment is withdrawn, suggesting that they may be at least operationally cured of their disease. Accurate definition of deep molecular responses (MRs) is therefore increasingly important for optimal patient management and comparison of independent data sets. We previously published proposals for broad standardized definitions of MR at different levels of sensitivity. Here we present detailed laboratory recommendations, developed as part of the European Treatment and Outcome Study for CML (EUTOS), to enable testing laboratories to score MR in a reproducible manner for CML patients expressing the most common BCR-ABL1 variants
Reference materials (RMs) for analysis of the human factor II (prothrombin) gene G20210A mutation
No full textThe Scientific Committee of Molecular Biology Techniques (C-MbT) in Clinical Chemistry of the IFCC has initiated a joint project in co-operation with the European Commission, Joint Research Centre, Institute of Reference Materials and Measurements to develop and produce plasmid-type reference materials (RMs), for the analysis of the human prothrombin gene G20210A mutation. Although DNA tests have a high impact on clinical decision-making and the number of tests performed in diagnostic laboratories is high, issues of quality and quality assurance exist, and currently only a few RMs for clinical genetic testing are available. A gene fragment chosen was produced that spans all primer annealing sites published to date. Both the wild-type and mutant alleles of this gene fragment were cloned into a pUC18 plasmid and two plasmid RMs were produced. In addition, a mixture of both plasmids was produced to mimic the heterozygous genotype. The present study describes the performance of these reference materials in a commutability study, in which they were tested by nine different methods in 13 expert laboratories.. This series of plasmid RMs are, to the best of our knowledge, the first plasmid-type clinical genetic RMs introduced worldwide
Rapid and highly specific screening for NPM1 mutations in acute myeloid leukemia
No full textNPM1 mutations, the most frequent molecular alterations in acute myeloid leukemia (AML), have become important for risk stratification and treatment decisions for patients with normal karyotype AML. Rapid screening for NPM1 mutations should be available shortly after diagnosis. Several methods for detecting NPM1 mutations have been described, most of which are technically challenging and require additional laboratory equipment. We developed and validated an assay that allows specific, rapid, and simple screening for NPM1 mutations. FAST PCR spanning exons 8 to 12 of the NPM1 gene was performed on 284 diagnostic AML samples. PCR products were visualized on a 2 % agarose E-gel and verified by direct sequencing. The FAST PCR screening method showed a specificity and sensitivity of 100 %, i.e., all mutated cases were detected, and none of negative cases carried mutations. The limit of detection was at 5-10 % of mutant alleles. We conclude that the FAST PCR assay is a highly specific, rapid (less than 2 h), and sensitive screening method for the detection of NPM1 mutations. Moreover, this method is inexpensive and can easily be integrated in the routine molecular diagnostic work-up of established risk factors in AML using standard laboratory equipment
Response to tyrosine kinase hnhibitors in myeloproliferative neoplasia with 8p11 translocation and CEP110-FGFR1 rearrangement
No full textThis brief communication reports on a patient with an exceedingly rare "8p11 (eight-p-eleven) myeloproliferative syndrome" (EMS) with CEP110-FGFR1 rearrangement who responded to treatment with the multi-tyrosine kinase inhibitor (TKI) dasatinib. Dasatinib improved quality of life substantially by increasing blood counts and reducing the need for transfusions. This report demonstrates that the second-generation TKI may provide a therapeutic option for elderly and frail EMS patients who cannot be offered aggressive therapy, including allogeneic hematopoietic cell transplantatio
Molecular diagnostics of myeloproliferative neoplasms
No full textSince the discovery of the JAK2 V617F mutation in the majority of the myeloproliferative neoplasms
(MPN) of polycythemia vera, essential thrombocythemia and primary myelofibrosis ten years ago, further
MPN-specific mutational events, notably in JAK2 exon 12, MPL exon 10 and CALR exon 9 have been
identified. These discoveries have been rapidly incorporated into evolving molecular diagnostic algorithms.
Whilst many of these mutations appear to have prognostic implications, establishing MPN diagnosis is of immediate clinical importance with selection, implementation and the continual evaluation of the
appropriate laboratory methodology to achieve this diagnosis similarly vital. The advantages and limitations of these approaches in identifying and quantitating the common MPN-associated mutations are considered
herein with particular regard to their clinical utility. The evolution of molecular diagnostic applications and
platforms has occurred in parallel with the discovery of MPN-associated mutations, and it therefore
appears likely that emerging technologies such as next-generation sequencing and digital PCR will in the
future play an increasing role in the molecular diagnosis of MPN.
Accepted for publication 30 April 2015 doi:10.1111/ejh.1257
Rapid and highly specific screening for NPM1 mutations in acute myeloid leukemia
No full textNPM1 mutations, the most frequent molecular alterations in acute myeloid leukemia (AML), have become important for risk stratification and treatment decisions for patients with normal karyotype AML. Rapid screening for NPM1 mutations should be available shortly after diagnosis. Several methods for detecting NPM1 mutations have been described, most of which are technically challenging and require additional laboratory equipment. We developed and validated an assay that allows specific, rapid, and simple screening for NPM1 mutations. FAST PCR spanning exons 8 to 12 of the NPM1 gene was performed on 284 diagnostic AML samples. PCR products were visualized on a 2% agarose E-gel and verified by direct sequencing. The FAST PCR screening method showed a specificity and sensitivity of 100%, i.e., all mutated cases were detected, and none of negative cases carried mutations. The limit of detection was at 5-10% of mutant alleles. We conclude that the FAST PCR assay is a highly specific, rapid (less than 2h), and sensitive screening method for the detection of NPM1 mutations. Moreover, this method is inexpensive and can easily be integrated in the routine molecular diagnostic work-up of established risk factors in AML using standard laboratory equipmen
Maximum power point tracking under realistic operating conditions
No full textThe process of tracking the Maximum Power Point (MPP), known as MPPT, becomes problematic under realistic operating conditions due to the potential for there to be more than one local maxima. A very detailed physics based model has been developed for a PV module (in this application a PV roof tile) using the Orcad platform for PSpice. This model is unusual in that it properly represents partial module shading and cell temperature variation. The PV roof tile, based on polycrystalline silicon cells, comprises 18 series-connected cells. In the model, each cell is represented by a standard two-diode sub-model, for which different levels of radiation and cell temperature can be simulated to obtain a realistic overall I-V characteristic for the module. The model can be extended to model any reasonable number of PV roof tiles wired in series and parallel to form a roof array. The IV characteristics calculated in this way using PSpice will be validated using an outdoor PV roof test system located at the University of Strathclyde, Glasgow
Impact of comorbidities on overall survival in patients with chronic myeloid leukemia: results of the randomized CML study IV.
No full textWe studied the influence of comorbidities on remission rate and overall survival (OS) in patients with chronic myeloid leukemia (CML). Participants of the CML Study IV, a randomized 5-arm trial designed to optimize imatinib therapy, were analyzed for comorbidities at diagnosis using the Charlson Comorbidity Index (CCI); 511 indexed comorbidities were reported in 1519 CML patients. Age was an additional risk factor in 863 patients. Resulting CCI scores were as follows: CCI 2, n = 589; CCI 3 or 4, n = 599; CCI 5 or 6, n = 229; and CCI ≥ 7, n = 102. No differences in cumulative incidences of accelerated phase, blast crisis, or remission rates were observed between patients in the different CCI groups. Higher CCI was significantly associated with lower OS probabilities. The 8-year OS probabilities were 93.6%, 89.4%, 77.6%, and 46.4% for patients with CCI 2, 3 to 4, 5 to 6, and ≥7, respectively. In multivariate analysis, CCI was the most powerful predictor of OS, which was still valid after removal of its age-related components. Comorbidities have no impact on treatment success but do have a negative effect on OS, indicating that survival of patients with CML is determined more by comorbidities than by CML itself. OS may therefore be inappropriate as an outcome measure for specific CML treatments. The trial was registered at www.clinicaltrials.gov as #NCT00055874
