Identification of Pseudoperonospora cubensis using real-time PCR and high resolution melting (HRM) analysis

2016 ◽  
Vol 82 (2) ◽  
pp. 110-115 ◽  
Author(s):  
Jeong Hwan Lee ◽  
Myong Ho Park ◽  
Sanghyeob Lee
Keyword(s):  
High Resolution ◽  
Real Time ◽  
Real Time Pcr ◽  
High Resolution Melting ◽  
Pseudoperonospora Cubensis ◽  
Hrm Analysis

scholarly journals Real-time PCR High-resolution Melting Analysis for the Species Identification of Meat Products: Focusing on Food Safety and Detection of Meat Adulterations

Thrita ◽  
2021 ◽  
Vol In Press (In Press) ◽  
Author(s):  
Peyman Gholamnezhad ◽  
Hamed Ahari ◽  
Gholamreza Nikbakht Brujeni ◽  
Seyed Amir Ali Anvar ◽  
Abbas Ali Motalebi
Keyword(s):  
High Resolution ◽  
Real Time ◽  
Species Identification ◽  
Real Time Pcr ◽  
High Resolution Melting ◽  
Meat Products ◽  
Hrm Analysis ◽  
Melting Analysis ◽  
Meat Species ◽  
Meat Samples

Background: Real-time polymerase chain reaction (PCR) and high-resolution melting (HRM) analysis are currently considered as reliable techniques for the species identification of meat-based products and widely used to detect meat adulteration. Objectives: To examine the validity of real-time PCR and HRM analysis to identify meat species in meat-based products. Methods: Meat samples from five species (i.e., cattle, sheep, chicken, turkey, and wild pig) were purchased. Minced meat from the animal species of interest was prepared at the purities of 10%, and 20% and also were prepared as single and mixtures of two species. For molecular assessments, DNA samples were extracted from all the meat samples and subjected to real-time PCR by amplifying a mitochondrial cytochrome b specific for each species. Results: All the meat species studied in this research were successfully detected in the mixed meat samples when separately examined by real-time PCR. High-resolution melting analysis showed that all the meat species of interest were efficiently distinguished when examined simultaneously. Conclusions: The data presented here shows that the real-time PCR and HRM analysis are reliable methods for the identification of meat species used in meat products.

scholarly journals High resolution melting analysis for the differentiation of Mycobacterium species

2014 ◽  
Vol 63 (10) ◽  
pp. 1284-1287 ◽  
Author(s):  
Rahizan Issa ◽  
Hatijah Abdul ◽  
Siti Hasmah Hashim ◽  
Valentinus H. Seradja ◽  
Nurul ‘Aishah Shaili ◽  
...  
Keyword(s):  
High Resolution ◽  
16S Rrna ◽  
Real Time ◽  
Real Time Pcr ◽  
High Resolution Melting ◽  
Rrna Gene ◽  
Hrm Analysis ◽  
Suitable Target ◽  
Effective Diagnosis ◽  
Dna Template

A quantitative real-time PCR (qPCR) followed by high resolution melting (HRM) analysis was developed for the differentiation of Mycobacterium species. Rapid differentiation of Mycobacterium species is necessary for the effective diagnosis and management of tuberculosis. In this study, the 16S rRNA gene was tested as the target since this has been identified as a suitable target for the identification of mycobacteria species. During the temperature gradient and primer optimization process, the melting peak (Tm) analysis was determined at a concentration of 50 ng DNA template and 0.3, 0.4 and 0.5 µM primer. The qPCR assay for the detection of other mycobacterial species was done at the Tm and primer concentration of 62 °C and 0.4 µM, respectively. The HRM analysis generated cluster patterns that were specific and sensitive to distinguished small sequence differences of the Mycobacterium species. This study suggests that the 16S rRNA-based real-time PCR followed by HRM analysis produced unique cluster patterns for species of Mycobacterium and could differentiate the closely related mycobacteria species.

scholarly journals Evaluation of High Resolution Melting (HRM) Analysis for Meat Species Identification of Raw and Cooked Meat

Separations ◽  
2021 ◽  
Vol 8 (8) ◽  
pp. 116
Author(s):  
Peyman Gholamnezhad ◽  
Hamed Ahari ◽  
Gholamreza Nikbakht Brujeni ◽  
Seyed Amir Ali Anvar ◽  
Abbasali Motallebi
Keyword(s):  
High Resolution ◽  
Real Time ◽  
Species Identification ◽  
Real Time Pcr ◽  
High Resolution Melting ◽  
Meat Products ◽  
Pcr Assay ◽  
Hrm Analysis ◽  
Mitochondrial Cytochrome B ◽  
Meat Samples

The current study aimed to examine a real-time PCR assay with high-resolution melting (HRM) analysis for the species identification of minced meat samples. Meat samples from several animal species were purchased and minced separately or as a mixture of two species. DNA was extracted from all meat samples and subjected to real-time PCR assay by amplifying species-specific mitochondrial cytochrome b regions. Regarding the meat mixtures, two separate melting curves with specific melt peak temperatures (Tm) were detected. Additionally, DNA from each species was quantified, based on the calibration curves. The results showed that a real-time PCR assay with HRM analysis is suitable for the species identification of meat products, and could be used for the detection of meat frauds.

scholarly journals Rapid and specific detection of porcine parvovirus using real-time PCR and High Resolution Melting (HRM) analysis

2015 ◽  
Vol 11 (1) ◽  
pp. 46 ◽  
Author(s):  
Hai-Qiong Yu ◽  
Xian-Quan Cai ◽  
Zhi-Xiong Lin ◽  
Xiang-Li Li ◽  
Qiao-Yun Yue ◽  
...  
Keyword(s):  
High Resolution ◽  
Real Time ◽  
Real Time Pcr ◽  
High Resolution Melting ◽  
Porcine Parvovirus ◽  
Specific Detection ◽  
Hrm Analysis

scholarly journals Mutation Scanning in a Single and a Stacked Genetically Modified (GM) Event by Real-Time PCR and High Resolution Melting (HRM) Analysis

2014 ◽  
Vol 15 (11) ◽  
pp. 19898-19923 ◽  
Author(s):  
Sina-Elisabeth Ben Ali ◽  
Zita Madi ◽  
Rupert Hochegger ◽  
David Quist ◽  
Bernhard Prewein ◽  
...  
Keyword(s):  
High Resolution ◽  
Real Time ◽  
Real Time Pcr ◽  
High Resolution Melting ◽  
Genetically Modified ◽  
Mutation Scanning ◽  
Hrm Analysis
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