Establishment of Cloning and Sequencing Method for High-Resolution HLA-B Genotype Assay

2014 ◽  
Vol 42 (11) ◽  
pp. 1574-1579 ◽  
Author(s):  
Xiao-Qing XING ◽  
Ya-Nan CHU ◽  
Zheng XIANG ◽  
Qin-Xin SONG ◽  
Guo-Hua ZHOU
Keyword(s):  
High Resolution ◽  
Cloning And Sequencing ◽  
Genotype Assay ◽  
Sequencing Method

scholarly journals High Resolution Melting Analysis in Leishmania Resistance: The Role of Multi Drug Resistance 1 Gene

2020 ◽  
Author(s):  
Maryam Fekri Soofi Abadi ◽  
Alireza Moradabadi ◽  
Reza Vahidi ◽  
Iman Rad ◽  
Shahriar Dabiri
Keyword(s):  
Drug Resistance ◽  
High Resolution ◽  
Genomic Dna ◽  
High Resolution Melting ◽  
Mdr1 Gene ◽  
Multi Drug Resistance ◽  
Biopsy Specimens ◽  
Sequencing Method ◽  
Melting Analysis

Abstract Objective Pentavalent antimonial compounds are currently used to treat Leishmaniasis. Resistance to these drugs is a serious problem. The purpose of this study was to evaluate the mutations in the multi-drug resistance 1 (MDR1) gene in biopsy specimens of Leishmania Tropica with high resolution melting (HRM) method.Results In this experimental study, genomic DNA was extracted from 130 patients with skin Leishmaniasis. Then, nucleotide changes were investigated throughout the gene length using HRM and sequencing methods. The results of the nucleotide changes showed that 61% of the samples that were unresponsive to drug had mutations in the MDR1 gene in different groups, which were confirmed by the sequencing method. These mutations can be one of the factors responsible for non-response to the treatment of the disease. HRM method can be used to diagnose drug resistance in Leishmaniasis. It is also recommended that further studies be done regarding the importance of drug resistance in the affected patients.

scholarly journals High resolution melting analysis and detection of Leishmania resistance: the role of multi drug resistance 1 gene

2021 ◽  
Vol 43 (1) ◽  
Author(s):  
Maryam Fekri Soofi Abadi ◽  
Alireza Moradabadi ◽  
Reza Vahidi ◽  
Saeedeh Shojaeepour ◽  
Sara Rostami ◽  
...  
Keyword(s):  
Drug Resistance ◽  
High Resolution ◽  
High Resolution Melting ◽  
Efflux Pump ◽  
Mdr1 Gene ◽  
Multi Drug Resistance ◽  
Sequencing Method ◽  
Melting Analysis ◽  
Resistance Protein

Abstract Background Pentavalent antimonial compounds are currently used to treat leishmaniasis and resistance to these drugs is a serious problem. Multidrug resistance protein is an efflux pump of the cell membrane that expels foreign compounds. This study designed to evaluate the mutations in the multi-drug resistance 1 (MDR1) gene, in biopsy specimens of Leishmania tropica, with high resolution melting (HRM) method. In this experimental study, genomic DNA was extracted from 130 patients with skin leishmaniasis. Then, nucleotide changes were investigated throughout the gene using HRM and sequencing methods. The samples categorized in 5 groups by differences in the melting temperature (Tm). Result The nucleotide changes analysis showed that 61% of the samples of different groups that were unresponsive to drug had mutations in the MDR1 gene, which were also confirmed by the sequencing method. These mutations can be one of the factors responsible for non-responsiveness to the treatment. Conclusion According to the findings, it seems that mutation in MDR1 gene could be responsible for drug resistance to pentavalent antimonial compounds. Furthermore, HRM method can be used to diagnose drug resistance in leishmaniasis. It is also recommended that further studies be done regarding the importance of drug resistance in the leishmania affected patients.

scholarly journals Determining spa-type of methicillin-resistant Staphylococcus aureus (MRSA) via High-Resolution Melting (HRM) analysis, Shiraz, Iran

2019 ◽  
Author(s):  
Zahra Hashemizadeh ◽  
Abdollah Bazargani ◽  
Samane Mohebi ◽  
Davood Kalantar-Neyestanaki ◽  
Nahal Hadi
Keyword(s):  
High Resolution ◽  
High Resolution Melting ◽  
Mutation Screening ◽  
Meca Gene ◽  
Gc Content ◽  
Teaching Hospitals ◽  
Spa Typing ◽  
Hrm Analysis ◽  
Sequencing Method ◽  
Spa Types

Abstract Objectives: Molecular typing methods are useful for rapid detection and control of a disease. Recently, the use of High-resolution melting (HRM) for spa typing of MRSA isolates were reported. This technique is rapid, inexpensive and simple for genotyping and mutation screening in DNA sequence. The aim of this study was to evaluate the ability of HRM-PCR to analysis spa genes amongst MRSA isolates.Results: A total of 50 MRSA isolates were collected from two teaching hospitals in Shiraz, Iran. The isolates were confirmed as MRSA by susceptibility to Cefoxitin and detection of mecA gene using PCR. we used High-resolution melting (HRM) analysis and PCR-sequencing method for spa typing of MRSA isolates. In total, 15 different spa types were discriminate by HRM and sequencing method. The melting temperature of the 15 spa types, using HRM genotyping were between 82.16° C and 85.66° C. The rate of GC% content was 39.4-46.3. According to the results, spa typing of 50 clinical isolates via PCR-sequencing and HRM methods were 100% similar. Consequently, HRM method can easily identify and rapidly differentiate alleles of spa genes. This method is faster, less laborious and more suitable for high sample at lower cost and risk of contamination.

scholarly journals High-Resolution Characterization of Toxoplasma gondii Transcriptome with a Massive Parallel Sequencing Method

DNA Research ◽  
2010 ◽  
Vol 17 (4) ◽  
pp. 233-243 ◽  
Author(s):  
J. Yamagishi ◽  
H. Wakaguri ◽  
A. Ueno ◽  
Y.-K. Goo ◽  
M. Tolba ◽  
...  
Keyword(s):  
High Resolution ◽  
Toxoplasma Gondii ◽  
Massive Parallel Sequencing ◽  
Parallel Sequencing ◽  
Sequencing Method

scholarly journals One-Step Differential Detection of OXA-48-Like Variants Using High-Resolution Melting (HRM) Analysis

Antibiotics ◽  
2020 ◽  
Vol 9 (5) ◽  
pp. 256
Author(s):  
Min Yi Lau ◽  
Kartini Abdul Jabar ◽  
Kek Heng Chua ◽  
Boon Pin Kee ◽  
Sasheela Sri La Sri Ponnampalavanar ◽  
...  
Keyword(s):  
High Resolution ◽  
High Resolution Melting ◽  
Bacterial Strains ◽  
Hrm Analysis ◽  
Sequencing Method ◽  
Tertiary Teaching ◽  
Carbapenemase Gene ◽  
Tertiary Teaching Hospital ◽  
One Step ◽  
Pcr Method

OXA-48-like carbapenemase gene remains a hidden threat, as different OXA-48 variants have varying presentations of susceptibility to antibiotics that might affect the treatment decisions. Rapid detection and differentiation of OXA-48-like carbapenemase genes are critical for targeted treatment and infection control. In this study, we aimed to develop high-resolution melting (HRM) analysis for the differentiation of OXA-48 variants. HRM analysis is a post-polymerase chain reaction (post-PCR) method for identification of small variations in nucleic acid sequences based on the PCR dissociation curve. A total of 82 bacterial strains, which consisted of Enterobacteriaceae and non-Enterobacteriaceae, were collected from a tertiary teaching hospital. The sensitivity and specificity of the assay were determined, and the developed assay was evaluated using the collected isolates against conventional-sequencing method. Overall, the developed assay was able to detect isolates that harboured OXA-48 and OXA232/OXA-181 by showing two distinct peaks at 81.1 ± 0.2 °C and 82.1 ± 0.2 °C, respectively. The detection limit of the assay was 1.6 x 10-6 ng/µl for OXA-48 and 1.8 x 10-7 ng/µl for OXA-232/OXA-181. This assay showed 100% specificity when evaluated on a panel of 37 isolates comprised of different species of bacteria and yeasts. When the assay with isolates collected in the year 2016 was first evaluated, the assay showed comparable results with conventional PCR-sequencing method where 34 OXA-48 and OXA-232/OXA-181 were detected. By using HRM analysis, the presence of OXA-48-like variants could be easily identified within 3 hours from the pure culture.

scholarly journals Use of the HRM Method in Quick Identification of FecXO Mutation in Highly Prolific Olkuska Sheep

Animals ◽  
2020 ◽  
Vol 10 (5) ◽  
pp. 844
Author(s):  
Grzegorz Smołucha ◽  
Katarzyna Piórkowska ◽  
Katarzyna Ropka-Molik ◽  
Jacek Sikora
Keyword(s):  
High Resolution ◽  
Molecular Biology ◽  
Sanger Sequencing ◽  
Genetic Basis ◽  
High Resolution Melting ◽  
Sheep Breed ◽  
Causative Mutation ◽  
Sequencing Method

Olkuska is a highly prolific sheep breed in Poland. Thanks to earlier identification of the genetic basis of its prolificacy, a mutation in the BMP-15 gene, we can use molecular biology tools to identify this causative mutation affecting prolificacy. In our research, we used the High-Resolution Melting (HRM) and Sanger sequencing methods to identify the genotypes of the studied animals. The result obtained by the HRM method is identical to those obtained by the sequencing method, which confirms the effectiveness of the HRM method and the possibility of quick and cheap identification of individuals with a FecXO mutation.

Observations of the spatial structure of interstellar hydrogen

1967 ◽  
Vol 31 ◽  
pp. 45-46
Author(s):  
Carl Heiles
Keyword(s):  
High Resolution ◽  
Spatial Structure ◽  
Velocity Dispersion ◽  
Temperature Variations

High-resolution 21-cm line observations in a region aroundlII= 120°,b11= +15°, have revealed four types of structure in the interstellar hydrogen: a smooth background, large sheets of density 2 atoms cm-3, clouds occurring mostly in groups, and ‘Cloudlets’ of a few solar masses and a few parsecs in size; the velocity dispersion in the Cloudlets is only 1 km/sec. Strong temperature variations in the gas are in evidence.

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