title: Rapid Identification of Echinococcus granulosus and E. canadensis Using High-Resolution Melting (HRM) Analysis by Focusing on a Single Nucleotide Polymorphism.
abstract: High-resolution melting (HRM) is a reliable and sensitive scanning method to detect variation in DNA sequences. We used this method to better understand the epidemiology and transmission of Echinococcus granulosus. We tested the use of HRM to discriminate the genotypes of E. granulosus and E. canadensis. One hundred forty-one hydatid cysts were collected from slaughtered animals in different parts of Isfahan-Iran in 2013. After DNA extraction, the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene was amplified using PCR coupled with the HRM curve. The result of HRM analysis using partial the sequences of cox1 gene revealed that 93, 35, and 2 isolates were identified as G1, G3, and G6 genotypes, respectively. A single nucleotide polymorphism (SNP) was found in locus 9867 of the cox1 gene. This is a critical locus for the differentiation between the G6 and G7 genotypes. In the phylogenic tree, the sample with a SNP was located between the G6 and G7 genotypes, which suggest that this isolate has a G6/G7 genotype. The HRM analysis developed in the present study provides a powerful technique for molecular and epidemiological studies on echinococcosis in humans and animals.
authors.affiliation: Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences.
authors.forename: Ahmad Hosseini
authors.initials: AH
authors.lastname: Safa
authors.forename: Majid Fasihi
authors.initials: MF
authors.lastname: Harandi
authors.forename: Mohammadhasan
authors.initials: M
authors.lastname: Tajaddini
authors.forename: Mohammad
authors.initials: M
authors.lastname: Rostami-Nejad
authors.forename: Mehdi
authors.initials: M
authors.lastname: Mohtashami-Pour
authors.forename: Nader
authors.initials: N
authors.lastname: Pestehchian
mesh_terms: D000818:Animals; D001681:Biological Assay / Q000662:veterinary*; D002417:Cattle; D018509:DNA, Helminth / Q000235:genetics* / Q000302:isolation & purification; D004443:Echinococcosis / Q000175:diagnosis / Q000453:epidemiology / Q000469:parasitology / Q000662:veterinary*; D004446:Echinococcus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D048209:Echinococcus granulosus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D003576:Electron Transport Complex IV / Q000235:genetics; D056426:Genetic Loci; D005838:Genotype; D006041:Goats; D015801:Helminth Proteins / Q000235:genetics*; D007492:Iran / Q000453:epidemiology; D009691:Nucleic Acid Denaturation; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide; D012756:Sheep
chemical_list: D018509:DNA, Helminth; D015801:Helminth Proteins; D003576:Electron Transport Complex IV