scholarly journals High-Resolution Melting Analysis as an Appropriate Method to Differentiate between Fasciola hepatica and F. gigantica

2019 ◽  
Author(s):  
Ahmad HOSSEINI-SAFA ◽  
Mohammad Bagher ROKNI ◽  
Sayed Hussain MOSAWI ◽  
Peyman HEYDARIAN ◽  
Hakim AZIZI ◽  
...  
Keyword(s):  
High Resolution ◽  
Real Time ◽  
Real Time Pcr ◽  
High Resolution Melting ◽  
Fasciola Hepatica ◽  
Pcr Amplification ◽  
Epidemiological Survey ◽  
North West ◽  
Melting Analysis ◽  
Fasciola Spp

Background: Fasciolosis is a shared disease between humans and livestock caused by hepatic trematodes; Fasciola hepatica and F. gigantica. Differentiate between the two species of this genus is essential. High-Resolution Melting (HRM) Analysis represents a new approach to this issue. This method can be performed right after termination of Real-Time PCR. This technique has not been used for identification of adult F. hepatica and F. gigantica genotypes. The aim of this study was to determine Fasciola species by using HRM in isolates taken from Iran, respectively. Methods: Ninety-three Fasciola spp. samples were collected from infected slaughtered animals in different regions of Iran, including North West (Ardebil Province) and South East (Zahedan Province) during 2016. Genomic DNA from the samples was extracted using a DNA extraction kit and then after Real-Time PCR amplification, HRM was done. Results: Overall, 59 and 34 isolates were identified as F. hepatica and F. gigantica, respectively. The percentages of each species from animals were as follows: sheep (F. hepatica, 80.39% and F. gigantica, 19.61%), cattle (F. hepatica, 42.85% and F. gigantica, 57.15%). Conclusion: HRM technique developed in the present study is a powerful, rapid and sensitive technique for epidemiological survey and molecular identification between F. hepatica and F. gigantica.

scholarly journals Specific Detection of Fasciola hepatica and F. gigantica in Infected Domesticated Animals Using High-Resolution Melting Analysis (HRM)

2020 ◽  
Author(s):  
Zeinab MOGHADAMIZAD ◽  
Ahmad HOSSEINI-SAFA ◽  
Mehdi MOHEBALI ◽  
Peyman HEYDARIAN ◽  
Mojgan ARYAEIPOUR ◽  
...  
Keyword(s):  
High Resolution ◽  
High Resolution Melting ◽  
Fasciola Hepatica ◽  
High Resolution Melting Analysis ◽  
Universal Primers ◽  
Specific Primer ◽  
Universal Primer ◽  
Melting Analysis ◽  
Liver Flukes ◽  
Fasciola Spp

Background: It is difficult to make an exact morphological distinction between Fasciola hepatica and Fasciola gigantica. We used High Resolution Melting analysis (HRM) method to differentiate the F. hepatica species from F. gigantica in order to differentiate them. Methods: Overall, 80 adult liver flukes were collected from infected slaughtered animals including cattle, sheep and goats from Lorestan Province, western Iran from Sep 2015 to Aug 2017. Genomic DNA was extracted using commercial DNA extraction kit. The multilocus sequences of mDNA including COX1, COX3 and ND6 were amplified employing real-time PCR & HRM analysis. Specific and universal primer pairs were designed for differentiation Fasciola spp. Results: Universal primers cannot be used to distinguish between these two species, but in the contrary, specific primer pairs of each species could differentiate them properly. Molecular identification using specific primer pairs were consistent. Conclusion: HRM is a simple, fast and reliable method for detecting and differentiating F. hepatica from F. gigantica and can be used for diagnostic and epidemiological purposes.

Development of a quantitation approach for total human and male DNA based on real time PCR followed by high resolution melting analysis

2016 ◽  
Vol 37 (21) ◽  
pp. 2734-2741 ◽  
Author(s):  
Santiago Ginart ◽  
Mariela Caputo ◽  
Evguenia Alechine ◽  
Daniel Corach ◽  
Andrea Sala
Keyword(s):  
High Resolution ◽  
Real Time ◽  
Real Time Pcr ◽  
High Resolution Melting ◽  
High Resolution Melting Analysis ◽  
Melting Analysis

Real-time PCR followed by high-resolution melting analysis – a new robust approach to evaluate SCCmectyping of methicillin-resistantStaphylococcus aureus

2019 ◽  
Vol 14 (2) ◽  
pp. 155-164 ◽  
Author(s):  
Yasaman Sadeghi ◽  
Seyed Alireza Salami ◽  
Pegah Kananizadeh ◽  
Sayed-Hamidreza Mozhgani ◽  
Mohammad Reza Pourmand
Keyword(s):  
High Resolution ◽  
Real Time ◽  
Real Time Pcr ◽  
High Resolution Melting ◽  
High Resolution Melting Analysis ◽  
Robust Approach ◽  
Melting 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.

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