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

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.

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Rapid identification of invasive fungal species using sensitive universal primers-based PCR and restriction endonuclease digestions coupled with high-resolution melting analysis

Journal of Microbiology Immunology and Infection ◽

10.1016/j.jmii.2019.06.001 ◽

2019 ◽

Vol 52 (5) ◽

pp. 728-735 ◽

Cited By ~ 1

Author(s):

Ming-Horng Tsai ◽

Lee-Chung Lin ◽

Jen-Fu Hsu ◽

Mei-Yin Lai ◽

Hsuan-Rong Huang ◽

...

Keyword(s):

High Resolution ◽

Restriction Endonuclease ◽

High Resolution Melting ◽

Rapid Identification ◽

Fungal Species ◽

High Resolution Melting Analysis ◽

Universal Primers ◽

Melting Analysis

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

Iranian Journal of Public Health ◽

10.18502/ijph.v48i3.894 ◽

2019 ◽

Cited By ~ 1

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.

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DNA-based Identification of Peucedanum ostruthium by High-Resolution-Melting Analysis (HRM)

Planta Medica ◽

10.1055/s-0032-1307632 ◽

2012 ◽

Vol 78 (05) ◽

Author(s):

C Schmiderer ◽

C Franz ◽

J Novak

Keyword(s):

High Resolution ◽

High Resolution Melting ◽

High Resolution Melting Analysis ◽

Melting Analysis

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Parental mosaicism in Marfan and Ehlers–Danlos syndromes and related disorders

European Journal of Human Genetics ◽

10.1038/s41431-020-00797-3 ◽

2021 ◽

Author(s):

Bertrand Chesneau ◽

Aurélie Plancke ◽

Guillaume Rolland ◽

Nicolas Chassaing ◽

Christine Coubes ◽

...

Keyword(s):

High Resolution ◽

Marfan Syndrome ◽

High Resolution Melting ◽

De Novo ◽

Direct Sequencing ◽

Causal Variant ◽

Connective Tissue Disorder ◽

High Resolution Melting Analysis ◽

Aortic Diseases ◽

Melting Analysis

AbstractMarfan syndrome (MFS) is a heritable connective tissue disorder (HCTD) caused by pathogenic variants in FBN1 that frequently occur de novo. Although individuals with somatogonadal mosaicisms have been reported with respect to MFS and other HCTD, the overall frequency of parental mosaicism in this pathology is unknown. In an attempt to estimate this frequency, we reviewed all the 333 patients with a disease-causing variant in FBN1. We then used direct sequencing, combined with High Resolution Melting Analysis, to detect mosaicism in their parents, complemented by NGS when a mosaicism was objectivized. We found that (1) the number of apparently de novo events is much higher than the classically admitted number (around 50% of patients and not 25% as expected for FBN1) and (2) around 5% of the FBN1 disease-causing variants were not actually de novo as anticipated, but inherited in a context of somatogonadal mosaicisms revealed in parents from three families. High Resolution Melting Analysis and NGS were more efficient at detecting and evaluating the level of mosaicism compared to direct Sanger sequencing. We also investigated individuals with a causal variant in another gene identified through our “aortic diseases genes” NGS panel and report, for the first time, on an individual with a somatogonadal mosaicism in COL5A1. Our study shows that parental mosaicism is not that rare in Marfan syndrome and should be investigated with appropriate methods given its implications in patient’s management.

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