Fast and Reliable Differentiation of Eight Trichinella Species Using a High Resolution Melting Assay

Warning

This publication doesn't include Faculty of Sports Studies. It includes Faculty of Science. Official publication website can be found on muni.cz.
Authors

RESLOVÁ Nikol ŠKORPÍKOVÁ Lucie SLANÝ Michal POZIO Edoardo KAŠNÝ Martin

Year of publication 2017
Type Article in Periodical
Magazine / Source Scientific Reports
MU Faculty or unit

Faculty of Science

Citation
Web https://www.nature.com/articles/s41598-017-16329-x
Doi http://dx.doi.org/10.1038/s41598-017-16329-x
Field Zoology
Keywords qPCR-HRMA; Trichinella; species identification; COI; ESV
Attached files
Description High resolution melting analysis (HRMA) is a single-tube method, which can be carried out rapidly as an additional step following real-time quantitative PCR (qPCR). The method enables the differentiation of genetic variation (down to single nucleotide polymorphisms) in amplified DNA fragments without sequencing. HRMA has previously been adopted to determine variability in the amplified genes of a number of organisms. However, only one work to date has focused on pathogenic parasites–nematodes from the genus Trichinella. In this study, we employed a qPCR-HRMA assay specifically targeting two sequential gene fragments–cytochrome c oxidase subunit I (COI) and expansion segment V (ESV), in order to differentiate 37 single L1 muscle larvae samples of eight Trichinella species. We prove that qPCR-HRMA based on the mitochondrial COI gene allows differentiation between the sequences of PCR products of the same length. This simple, rapid and reliable method can be used to identify at the species level single larvae of eight Trichinella taxa.
Related projects:

You are running an old browser version. We recommend updating your browser to its latest version.

More info