GENOTYPING OF EIGHT TRICHINELLA SPECIES USING HIGH RESOLUTION MELTING ANALYSIS

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14310%2F17%3A00097650" target="_blank" >RIV/00216224:14310/17:00097650 - isvavai.cz</a>

Výsledek na webu

—

DOI - Digital Object Identifier

—

Jazyk výsledku

angličtina

Název v původním jazyce

GENOTYPING OF EIGHT TRICHINELLA SPECIES USING HIGH RESOLUTION MELTING ANALYSIS

Popis výsledku v původním jazyce

Nematodes of the genus Trichinella are worldwide distributed foodborne parasites with the high impact on public health and animal trade. They invade small intestine cells and skeletal muscles of wide range of mammals, birds and reptiles. Trichinella spp. are causative agents of human trichinellosis, a serious disease which has been documented in 55 countries with an average incidence of approximately ten-thousand cases per year. According to ability of muscle larvae to form a collagen capsule, within the genus Trichinella two main clades are recognized. The “encapsulated clade” contains six species (T1 - Trichinella spiralis, T2 - T. nativa, T3 - T. britovi, T5 - T. murrelli, T7 - T. nelsoni, T12 - T. patagoniensis) and three additional taxonomically undefined genotypes (T6, T8 and T9). “Non-encapsulated clade” includes three species (T4 - T. pseudospiralis, T10 - T. papuae and T11 - T. zimbabwensis). Despite the difference in capsule formation and some size differences, the species and genotypes of all developmental stages of these parasites are morphologically indistinguishably. Therefore, we have developed a high resolution melting analysis (HRMA) for the unequivocal differentiation of eight Trichinella species (T1, T2, T3, T4, T5, T7, T10 and T11). The assay was based on the mitochondrial cytochrome c oxidase subunit I (COI) gene, representing the variable part of DNA sequence which shows high sequence divergence even among closely allied species. The genomic DNA of a single muscle larva was used as a template in qPCR, the polymorphic region of the COI gene was amplified and the obtained amplicons were analysed for species-specific HRMA curves.

Název v anglickém jazyce

GENOTYPING OF EIGHT TRICHINELLA SPECIES USING HIGH RESOLUTION MELTING ANALYSIS

Popis výsledku anglicky

Nematodes of the genus Trichinella are worldwide distributed foodborne parasites with the high impact on public health and animal trade. They invade small intestine cells and skeletal muscles of wide range of mammals, birds and reptiles. Trichinella spp. are causative agents of human trichinellosis, a serious disease which has been documented in 55 countries with an average incidence of approximately ten-thousand cases per year. According to ability of muscle larvae to form a collagen capsule, within the genus Trichinella two main clades are recognized. The “encapsulated clade” contains six species (T1 - Trichinella spiralis, T2 - T. nativa, T3 - T. britovi, T5 - T. murrelli, T7 - T. nelsoni, T12 - T. patagoniensis) and three additional taxonomically undefined genotypes (T6, T8 and T9). “Non-encapsulated clade” includes three species (T4 - T. pseudospiralis, T10 - T. papuae and T11 - T. zimbabwensis). Despite the difference in capsule formation and some size differences, the species and genotypes of all developmental stages of these parasites are morphologically indistinguishably. Therefore, we have developed a high resolution melting analysis (HRMA) for the unequivocal differentiation of eight Trichinella species (T1, T2, T3, T4, T5, T7, T10 and T11). The assay was based on the mitochondrial cytochrome c oxidase subunit I (COI) gene, representing the variable part of DNA sequence which shows high sequence divergence even among closely allied species. The genomic DNA of a single muscle larva was used as a template in qPCR, the polymorphic region of the COI gene was amplified and the obtained amplicons were analysed for species-specific HRMA curves.

Druh

O - Ostatní výsledky

CEP obor

—

OECD FORD obor

10600 - Biological sciences

Projekt

—

Návaznosti

S - Specificky vyzkum na vysokych skolach

Rok uplatnění

2017

Kód důvěrnosti údajů

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