Identification of oncogene mutations in leukemia patients using microchip-based PCR analysis

Богданов, Константин; Nikulina, Tatiana; Lomaia, Elza; Slyadnev, М. N.; Zaritskey, Andrey

doi:10.1134/s1068162017040033

Cited by 6 publications

(3 citation statements)

References 17 publications

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“…However, in our case, this assumption may be true only in part. Thus, a reduced number (2-3, no multiplication by 100) of NPM1/A gene copies was found in 4 of 6 patients with a low (<0.5) allelic load of the FLT3-ITD mutation, whereas only 3 of 8 patients with a high (>0.5) allelic load of FLT3-ITD had an elevated number (11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22)(23)(24)(25) of NPM1/A gene copies (data not shown). Thus, it is still unclear which additional causes may contribute to a worse prognosis in patients carrying the FLT3-ITD mutation.…”

Section: Discussionmentioning

confidence: 93%

“…To detect various oncogene mutations ( AML1-ETO , CBFβ-MYH11 , PML-RARα , BCR-ABL , DEK-NUP214 , NUP98-NSD1 , MLL-AF4 ), cDNA was amplified using microarray real-time PCR after reverse transcription (RT) [ 23 ]. Each sample for PCR contained 1× PCR buffer, 2.5 mM of each dNTP, 10 pM forward and reverse primers, 6 pM FAM-labeled probe at the 5′-end, 10 ng of cDNA, and 5 units of Taq polymerase (Evrogen, Moscow, Russia).…”

Section: Methodsmentioning

confidence: 99%

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Shift of N-MYC Oncogene Expression in AML Patients Carrying the FLT3-ITD Mutation

et al. 2023

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Mutations in the FLT3 gene not only lead to abnormalities in its structure and function, but also affect the expression of other genes involved in leukemogenesis. This study evaluated the expression of genes that are more characteristic of neuroblastoma but less studied in leukemia. N-MYC oncogene expression was found to be more than 3-fold higher in primary AML patients carrying the FLT3-ITD mutation compared to carriers of other mutations as well as patients with normal karyotype (p = 0.03946). In contrast to the expression of several genes (C-MYC, SPT16, AURKA, AURKB) directly correlated to the allelic load of FLT3-ITD, the expression of the N-MYC oncogene is extremely weakly related or independent of it (p = 0.0405). Monitoring of N-MYC expression in some patients with high FLT3-ITD allelic load receiving therapy showed that a decrease in FLT3-ITD allelic load is not always accompanied by a decrease in N-MYC expression. On the contrary, N-MYC expression may remain elevated during the first three months after therapy, which is additional evidence of the emergence of resistance to therapy and progression of AML.

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Section: Discussionmentioning

confidence: 93%

Section: Methodsmentioning

confidence: 99%

Shift of N-MYC Oncogene Expression in AML Patients Carrying the FLT3-ITD Mutation

et al. 2023

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“…The microchip consists of an aluminum metal plate with stamped microwells which provides high heat transfer efficiency and offers a high surface area to volume ratio with a 1.2 μl reaction well [5]. These attributes of product design combine to give high heating and cooling rates, low reagent consumption, rapid results, and are overall cost-effective [7]- [12]. These attributes also allow this system to be a candidate to become an effective workhorse in the current or post-pandemic scenario.…”

Section: Introductionmentioning

confidence: 99%

Validation of Microchip RT-PCR COVID-19 Detection System

Campos-Stairiker¹,

Pidathala²,

Gill³

et al. 2021

JBM

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While meeting the pandemic demand of SARS-CoV-2 testing, clinical laboratories worldwide tend to adopt new test systems offering cost-effective and faster test outcomes. However, the reliability of SARS-CoV-2 test results has paramount importance in the management of such a health crisis. Therefore, this study sought to determine the accuracy of the test results from a novel duplex Microchip RT-PCR test system using patient saliva samples and nasal swabs stabilized in Viral Transport Medium (VTM) with reference threshold Cycle Values (Ct). The VTM used to stabilize these samples during transport was found to be inhibitory to the RT-PCR. Therefore, all the samples were subjected to spin column purification of total RNA to remove the influence of VTM. A total of 70 patient samples, including 24 positive-and 31 negative-saliva in VTM samples and 15 positive nasal swab samples, were tested. Results obtained from both the sample types were compared to their reference values and no false positive or false negatives were observed. From this data, accuracy, specificity, and sensitivity were determined to be 100% applying the corresponding formulae. The limit of detection with 95% confidence probability was determined to be 2.5 copies/µl in the original sample.

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Microchip RT-PCR Detection of Nasopharyngeal SARS-CoV-2 Samples

Cojocaru

Yaseen

Unrau

et al. 2021

The Journal of Molecular Diagnostics

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Fast, accurate and reliable diagnostic tests are critical for controlling the spread of the 2019 Coronavirus disease associated with SARS-CoV-2 infection. The current gold standard for testing is real time PCR (RT-PCR), however, during the current pandemic, supplies of testing kits and reagents have been limited. We report the validation of a rapid (30 min), user-friendly and accurate microchip RT-PCR assay for detection of SARS-CoV-2 from nasopharyngeal swab RNA extracts. Microchips pre-loaded with COVID-19 primers and probes for the N gene accommodate 1.2 μl reaction volumes, lowering the required reagents by 10-fold compared to tube-based RT-PCR. We validated our assay using contrived reference samples and 21 clinical samples from patients in Canada, determining a limit of detection of 1 copy per reaction. The microchip RT-PCR provides a significantly lower resource alternative to the CDC approved real-time RT-PCR assays with comparable sensitivity, showing 100% positive and negative predictive agreement of clinical samples.

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Identification of oncogene mutations in leukemia patients using microchip-based PCR analysis

Cited by 6 publications

References 17 publications

Shift of N-MYC Oncogene Expression in AML Patients Carrying the FLT3-ITD Mutation

Shift of N-MYC Oncogene Expression in AML Patients Carrying the FLT3-ITD Mutation

Validation of Microchip RT-PCR COVID-19 Detection System

Microchip RT-PCR Detection of Nasopharyngeal SARS-CoV-2 Samples

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