Format

Send to

Choose Destination
Parasitol Res. 2016 Jul;115(7):2715-20. doi: 10.1007/s00436-016-5018-1. Epub 2016 Mar 23.

High-resolution melting analysis (HRM) for differentiation of four major Taeniidae species in dogs Taenia hydatigena, Taenia multiceps, Taenia ovis, and Echinococcus granulosus sensu stricto.

Author information

1
Research Center for Hydatid Disease in Iran, Department of Parasitology, Kerman University of Medical Sciences, Kerman, 7616914115, Iran.
2
Department of Research and Technology, Alborz University of Medical Sciences, Karaj, Iran.
3
School of Medicine, Shahroud University of Medical Sciences, Shahroud, Iran.
4
Research Center for Hydatid Disease in Iran, Department of Parasitology, Kerman University of Medical Sciences, Kerman, 7616914115, Iran. majid.fasihi@gmail.com.

Abstract

Tapeworms of the genus Taenia include several species of important parasites with considerable medical and veterinary significance. Accurate identification of these species in dogs is the prerequisite of any prevention and control program. Here, we have applied an efficient method for differentiating four major Taeniid species in dogs, i.e., Taenia hydatigena, T. multiceps, T. ovis, and Echinococcus granulosus sensu stricto. High-resolution melting (HRM) analysis is simpler, less expensive, and faster technique than conventional DNA-based assays and enables us to detect PCR amplicons in a closed system. Metacestode samples were collected from local abattoirs from sheep. All the isolates had already been identified by PCR-sequencing, and their sequence data were deposited in the GenBank. Real-time PCR coupled with HRM analysis targeting mitochondrial cox1 and ITS1 genes was used to differentiate taeniid species. Distinct melting curves were obtained from ITS1 region enabling accurate differentiation of three Taenia species and E. granulosus in dogs. The HRM curves of Taenia species and E .granulosus were clearly separated at Tm of 85 to 87 °C. In addition, double-pick melting curves were produced in mixed infections. Cox1 melting curves were not decisive enough to distinguish four taeniids. In this work, the efficiency of HRM analysis to differentiate four major taeniid species in dogs has been demonstrated using ITS1 gene.

KEYWORDS:

Echinococcus granulosus; HRM analysis; ITS1; Melting curve; Taenia; Taeniidae

PMID:
27008188
DOI:
10.1007/s00436-016-5018-1
[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for Springer
Loading ...
Support Center