High- Resolution Melting (Hrm) Curve Analysis: New Approach Used to Detect Blad and Dumps in Uruguayan Holstein Breed

Federici, María Teresa; Sica, Andrea Branda; Artigas, Rody; Briano, Carolina; Dutra, Fernando; Llambí, Silvia

doi:10.5380/avs.v23i4.62578

Cited by 4 publications

(2 citation statements)

References 9 publications

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“…In addition to ease and cost-effectiveness, this method is capable of detecting both Table 3. Demographic Information homozygote and heterozygote types of mutations, unlike some other PCR-based methods (Moradabadi et al, 2017;Federici et al, 2018;Er and Chang, 2012;Montgomery et al, 2010;Norambuena et al, 2009). The prevalence of N-RAS hot-spot mutations was 14% in adult AML patients in our study.…”

Section: Discussionmentioning

confidence: 50%

“…Thereupon, in the HRM step, the PCR products are gradually heated, and in the melting temperature, the DNA samples are being denatured, causing fluorescence fading. Finally, a melt curve is drawn according to the fluorescence level as opposed to the temperature that even a single base change will cause alterations in the resultant melting curve (Moradabadi et al, 2017;Federici et al, 2018;Er and Chang, 2012;Montgomery et al, 2010;Norambuena et al, 2009;Hassani et al, 2017;Moonesi et al, 2021).…”

Section: Rapid Detection Of N-ras Gene Common Mutations In Acutementioning

confidence: 99%

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Rapid Detection of N-RAS Gene Common Mutations in Acute Myeloid Leukemia (AML) Using High Resolution Melting (HRM) Method

Khosravi

Moonesi

Moradabadi

et al. 2022

Asian Pac J Cancer Prev

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Objective: Acute myeloid leukemia is caused by the clonal proliferation of undifferentiated myeloid hematopoietic precursors. AML prognosis is highly involved in the treatment response and is determined by mutations in several genes such as N-RAS. This study aims to identify the distribution of common N-RAS mutations (codons 12, 13, and 61) in AML patients using the HRM method and confirm this method's efficiency for mutation detection by comparing its results with the sequencing data as the Gold standard method. Methods: Peripheral blood samples were taken from 50 newly diagnosed AML patients. Mononuclear cells were isolated from samples, and DNA was extracted. Then, mutation detection was investigated using the HRM method. Efficacy of the HRM method in mutation detection was determined in comparison with direct sequencing. Results: N-RAS mutations were detected in 7 of the 50 samples (14%). Most of the mutations were found in codon 12 (57.14%), and 28.57% and 14.28% of mutations were in codons 61 and 13, respectively. There was no statistically significant association between patients' demographic data and HRM results. Conclusion: According to mutation detection results and the HRM results confirmation with the sequencing method, this method can be introduced as an efficient, low-cost, and fast method for detecting common mutations.

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Section: Discussionmentioning

confidence: 50%

Section: Rapid Detection Of N-ras Gene Common Mutations In Acutementioning

confidence: 99%

Rapid Detection of N-RAS Gene Common Mutations in Acute Myeloid Leukemia (AML) Using High Resolution Melting (HRM) Method

Khosravi

Moonesi

Moradabadi

et al. 2022

Asian Pac J Cancer Prev

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Detection of the Brachyspina mutation in Uruguayan Holstein cows using real time PCR and melting curve analysis

et al. 2021

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Brachyspina syndrome (BS) is a rare monogenic autosomal recessive hereditary disorder of the Holstein Fresian breed caused by a deletion of 3.3Kb in the Fanconi anemia complementation group I (FANCI) gene on BTA-21, which leads to a frame-shift and premature stop codon. Some of the consequences of BS are the reduction of the fertility rate and milk production. This study developed a simple, sensitive, rapid cost- effective assay method based on real time PCR and melting curve analysis for the detection of BS carrier animals. Sixty-eight normal homozygous and four heterozygous carrier genotypes were detected and confirmed through traditional PCR- electrophoresis analysis. We concluded that the assay we have developed proved to be a reliable, highly precise and low-cost tool, which could be used to monitor the presence of the BS mutation in uruguayan Holstein breed.

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The Effect of DNA Concentration on the HRM Performance in Detecting Jak2 p.V617F Variant in Patients with Myeloproliferative Neoplasms

Moradabadi

Soltani

Shariati

et al. 2023

CST

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Introduction: Janus kinase 2 (JAK2) is an intracellular signaling protein. JAK2 p. V617F is a common variant in normal karyotype myeloproliferative neoplasms (MPN). High-resolution melting (HRM) analysis is one of the essential methods for detecting the JAK2 p.V617F variant. In this study, we have investigated the effect of DNA concentration on detecting the JAK2 p.V617F variant using the HRM method. Materials and Method: Genomic DNA was extracted from human blood and diluted ten times in distilled water from 1 to 0.03; afterward, HRM was conducted for each dilution (triplicate). Using SPSS v.20.0 software, the mean Tm of each dilution was calculated and compared. Results: The HRM results revealed the JAK2 wild type and variant to have Tms of 81/64°C and 80/76°C, respectively. At the endpoint of the pre-amplification, the dilutions had different emissions. The statistical analysis revealed no statistically significant differences in Tm between samples with varying DNA concentrations (P value > 0.05). Conclusion: There have been no significant differences obtained in the analysis of JAK2 p.V617F point variant in different DNA dilutions, implying that the HRM analysis has no relation to DNA concentration.

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High- Resolution Melting (Hrm) Curve Analysis: New Approach Used to Detect Blad and Dumps in Uruguayan Holstein Breed

Cited by 4 publications

References 9 publications

Rapid Detection of N-RAS Gene Common Mutations in Acute Myeloid Leukemia (AML) Using High Resolution Melting (HRM) Method

Rapid Detection of N-RAS Gene Common Mutations in Acute Myeloid Leukemia (AML) Using High Resolution Melting (HRM) Method

Detection of the Brachyspina mutation in Uruguayan Holstein cows using real time PCR and melting curve analysis

The Effect of DNA Concentration on the HRM Performance in Detecting Jak2 p.V617F Variant in Patients with Myeloproliferative Neoplasms

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