Tag Archives: RT-PCR

Clear or White? A RT-PCR plate comparison for SARS-CoV-2 diagnosis

DOI: 10.2478/amma-2022-0024

Objective: During the COVID-19 pandemic, laboratories have used various extraction and amplification kits, associated with different auxiliary materials. This study aims to investigate how different types of plates may affect RT-PCR performance.
Methods: Data on the positive controls (PCs) of RT-PCR runs for SARS-CoV-2 detection between December 2020 and April 2022 was collected retrospectively in the Molecular biology department of the County Emergency Clinical Hospital of Târgu Mureș’s clinical laboratory. The materials used consisted in MOLgen SARS-CoV-2 (Adaltis) and EliGene COVID19 CONFIRM 500 R amplification kits, 96-well full-skirted white and clear plates, and clear films. Microsoft Excel was used for the database and it included information about Cycle threshold (Ct) and maximum fluorescence. Statistical analysis, performed in MedCalc, consisted of Grubbs test, Kolmogorov–Smirnov Test, F test, T student test, and Mann-Whitney test to compare central tendencies. The significance threshold was set at p<0.05.
Results: The Ct comparison for MOLgen kit white plates vs clear plates: FAM channel- U=1052.5, Z=2.07, p=0.038, medians for white plates and clear plates were 22.80 and 23.25, respectively; ROX channel- U=784, Z=3.21, p=0.001, medians 21.93 and 21.43, respectively; Cy5 channel- U=1028.5, Z=1.95, p=0.518, medians 22.12, 21.75, respectively. For EliGene kit: U=848.5, Z=3.27, p=0.001, medians 28.26 (white plates) and 28.0 (clear plates). Comparison of the maximum fluorescence reached on both kits with white and clear plates computed p values <0.0001.
Conclusions: Between white and clear plates there are statistically significant differences considering Ct values and maximum read fluorescence, but with no impact on test outcome.

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The Results of Allogeneic Stem Cell Transplantation in CML — the Experience of BMT Unit Tîrgu Mureş

Introduction: Chronic myeloid leukemia (CML) is a clonal myeloproliferative disorder the molecular hallmark of the disease is the BCR-ABL gene rearrangement which occurs as the result of a reciprocal translocation between chromosomes 9 and 22. Imatinib, a small molecule, tyrosine kinase inhibitor (TKI) was the first drug that targeted BCR-ABL. Since the introduction of the first and second generation of TKI the role of allogeneic stem cell transplantation in chronic myeloid leukemia is being reevaluated. With this retrospective analysis our aim was to define the role of allogeneic stem cell transplantation for CML in the tyrosine kinase inhibitor era. The following is a general overview of the role of ASCT in the management of CML.
Material and methods: At the BMT Unit Tîrgu Mureș between 2005–2009 we performed five allogeneic transplantations of high risk CML patients with identical sibling donors.
Results: Two of the patients are at present in complete hematologic and cytogenetic remission with no or minimal immunosuppressive therapy after 6 and respectively 3 years of follow up time. Two of the patients had disease free survival but died from infectious complications appeared in the 3rd and 6th month after the allogeneic stem cell transplantation. One patient had an early relapse with treatment refractory disease and died from the evolution of the disease.
Conclusions: We perform allogeneic stem cell transplantation only in the cases in which we have resistance to first and second generation of tyrosine kinase inhibitors (TKI), intolerance to TKI and if we have a suitable donor.

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