Differentiation of Eight Trichinella Species Using a High Resolution Melting Assay

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00027162%3A_____%2F18%3AN0000240" target="_blank" >RIV/00027162:_____/18:N0000240 - isvavai.cz</a>

Výsledek na webu

<a href="https://hd2018.natur.cuni.cz/programme" target="_blank" >https://hd2018.natur.cuni.cz/programme</a>

DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

Differentiation of Eight Trichinella Species Using a High Resolution Melting Assay

Popis výsledku v původním jazyce

24th Helminthological Days. Cosmopolitan nematodes of the genus Trichinella are foodborne parasites with the high impact on animal trade and food safety. A broad range of carnivore and omnivore animals have been reported to be hosts of these parasites, including the economically important domestic pig. The representatives of the genus Trichinella are also causative agents of human trichinel-losis, a serious human disease, which has been documented at least in 55 countries around the world. Based on zoogeographical, epidemiological and molecular characters, there are 12 taxa recognised in the genus Trichinella. According to ability of muscle larvae to encapsulate in host muscle tissues or not, two main clades (encapsulated and non-encapsulated) are recog-nized. Despite this difference in capsule formation, there are no clear morphological features useful for species differentiation. In this study, we adopted a high resolution melting analysis, a single-tube method, which can be carried out as an additional step following a real-time quantitative PCR. This method enables the distinguishing of genetic variation (down to single nucleotide polymorphisms) in amplified DNA fragments without necessity of subsequent se-quencing. Our approach differentiating capacity was evaluated using eight Trichinella species (T1, T2, T3, T4, T5, T7, T10 and T11), 37 samples of genomic DNA isolated from single muscle larvae were analyzed. The assay was based on polymorphisms in 240 bp long frag-ment of the cytochrome c oxidase subunit I gene. This mitochondrial gene shows high se-quence divergence even among closely allied species. For the evaluation of melting process have been used difference plot, which represents the most transparent form of expression of species-specific matrix curves. We show that high resolution melting analysis is rapid and reliable method that can be used to identify single larvae of eight Trichinella species. In the future we envisage that Trichinella species-specific matrix curves could be automatically evaluated using the computer application.

Název v anglickém jazyce

Differentiation of Eight Trichinella Species Using a High Resolution Melting Assay

Popis výsledku anglicky

24th Helminthological Days. Cosmopolitan nematodes of the genus Trichinella are foodborne parasites with the high impact on animal trade and food safety. A broad range of carnivore and omnivore animals have been reported to be hosts of these parasites, including the economically important domestic pig. The representatives of the genus Trichinella are also causative agents of human trichinel-losis, a serious human disease, which has been documented at least in 55 countries around the world. Based on zoogeographical, epidemiological and molecular characters, there are 12 taxa recognised in the genus Trichinella. According to ability of muscle larvae to encapsulate in host muscle tissues or not, two main clades (encapsulated and non-encapsulated) are recog-nized. Despite this difference in capsule formation, there are no clear morphological features useful for species differentiation. In this study, we adopted a high resolution melting analysis, a single-tube method, which can be carried out as an additional step following a real-time quantitative PCR. This method enables the distinguishing of genetic variation (down to single nucleotide polymorphisms) in amplified DNA fragments without necessity of subsequent se-quencing. Our approach differentiating capacity was evaluated using eight Trichinella species (T1, T2, T3, T4, T5, T7, T10 and T11), 37 samples of genomic DNA isolated from single muscle larvae were analyzed. The assay was based on polymorphisms in 240 bp long frag-ment of the cytochrome c oxidase subunit I gene. This mitochondrial gene shows high se-quence divergence even among closely allied species. For the evaluation of melting process have been used difference plot, which represents the most transparent form of expression of species-specific matrix curves. We show that high resolution melting analysis is rapid and reliable method that can be used to identify single larvae of eight Trichinella species. In the future we envisage that Trichinella species-specific matrix curves could be automatically evaluated using the computer application.

Druh

O - Ostatní výsledky

CEP obor

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OECD FORD obor

10608 - Biochemistry and molecular biology

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2018

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů