Genotyping anisakid nematodes using HRM analysis

3^rd International Conference on Genomics & Pharmacogenomics

September 21-23, 2015 San Antonio, USA

Cavallero S1, Umehara A2 and D�Amelio S1

1Sapienza University of Rome, Italy 2Azabu University, Japan

Posters-Accepted Abstracts: J Mol Genet Med

Abstract :

Anisakids are parasitic nematodes belonging to the super family Ascaridoidea. The presence of infective larvae in fish intended for human consumption cause economic and medical problems. Among anisakids, Anisakis simplex sensu lato and Pseudoterranova decipiens s. l are the species mainly responsible for human anisakidosis. Rarely, Contracaecum sp. and Hysterothylacium sp. (Raphidascaridae Family) have been found in association with accidental found in gastric/intestinal tract, even if the latter is commonly considered not pathogenic to humans. The aim of the study was to develop a new molecular approach to differentiate and identify anisakids based on the High Resolution Melting (HRM) of a selected molecular marker (nuclear ribosomal ITS). By now, only A. simplex complex was discriminated using HRM. Representative species of Anisakis, Pseudoterranova, Contracaecum and Hysterothylacium were tested to verify the presence of diagnostic profiles to differentiate nematodes at genus and species level. HRM profiles of Anisakis simplex s. l revealed Tm of 78.0-78.2 �°C for A. simplex s. s., of 79.4-79.5 �°C for A. pegreffii and of 78.4-78.6 �°C for hybrids. Members of P. decipiens complex were tested with Anisakis spp., Contracaecum osculatum and Hysterothylacium aduncum. Results revealed no amplification for all Anisakis spp and three HRM profiles: One showed Tm of 79.4-79.5 �°C for P. decipiens sensu stricto and P. krabbei; one showed 79.9-80.0 �°C for P. azarasi and P. cattani; one showed 79.1-79.2 �°C for C. osculatum and H. aduncum. Preliminary results obtained indicate the usefulness of such genotyping approach, potentially functional for screening in basic research area, food industry and diagnostics.

Biography :

Email: serena.cavallero@uniroma1.it