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High resolution melting technique for molecular epidemiological studies of cystic echinococcosis: Differentiating G1, G3, and G6 genotypes of Echinococcus granulosus sensu lato

Rostami, S. and Talebi, S. and Babaei, Z. and Sharbatkhori, M. and Ziaali, N. and Rostami, H. and Harandi, M.F. (2013) High resolution melting technique for molecular epidemiological studies of cystic echinococcosis: Differentiating G1, G3, and G6 genotypes of Echinococcus granulosus sensu lato. Parasitology Research, 112 (10). pp. 3441-3447. ISSN 09320113 (ISSN)

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Abstract

Reliable and rapid genotyping of large number of Echinococcus granulosus sensu lato isolates is crucial for understanding the epidemiology and transmission of cystic echinococcosis. We have developed a method for distinguishing and discriminating common genotypes of E. granulosus s.l. (G1, G3, and G6) in Iran. This method is based on polymerase chain reaction coupled with high resolution melting curve (HRM), ramping from 70 to 86 C with fluorescence data acquisition set at 0.1 C increments and continuous fluorescence monitoring. Consistency of this technique was assessed by inter- and intra-assays. Assessment of intra- and inter-assay variability showed low and acceptable coefficient of variations ranging from 0.09 to 0.17 %. Two hundred and eighty E. granulosus s.l. isolates from sheep, cattle, and camel were used to evaluate the applicability and accuracy of the method. The isolates were categorized as G1 (93, 94, and 25 %), G3 (7, 4, and 4 %), and G6 (0, 2, and 71 %) for sheep, cattle, and camel, respectively. HRM results were completely compatible with those obtained from sequencing and rostellar hook measurement. This method proved to be a valuable screening tool for large-scale molecular epidemiological studies. © 2013 Springer-Verlag Berlin Heidelberg.

Item Type: Article
Additional Information: Unmapped bibliographic data: LA - English [Field not mapped to EPrints] J2 - Parasitol. Res. [Field not mapped to EPrints] C2 - 23832641 [Field not mapped to EPrints] AD - Research Center of Tropical and Infectious Diseases, Kerman University of Medical Sciences, Kerman, Iran [Field not mapped to EPrints] AD - Department of Medical Parasitology, School of Medicine, Kerman University of Medical Sciences, Kerman 76169-14111, Iran [Field not mapped to EPrints] AD - Department of Medical Genetics, Tehran University of Medical Sciences, Tehran, Iran [Field not mapped to EPrints] AD - Infectious Diseases Research Center, Department of Medical Parasitology and Mycology, Golestan University of Medical Sciences, Gorgan, Iran [Field not mapped to EPrints] AD - Shariati Hospital, Karaj Alborz, Iran [Field not mapped to EPrints] AD - Research Center for Hydatid Disease in Iran, Kerman University of Medical Sciences, Kerman 76169-14111, Iran [Field not mapped to EPrints] DB - Scopus [Field not mapped to EPrints]
Uncontrolled Keywords: article, camel, cattle, disease transmission, DNA sequence, echinococcosis, Echinococcus granulosus, fluorescence, genotype, high resolution melting analysis, Iran, molecular epidemiology, nonhuman, nucleotide sequence, parasite isolation, polymerase chain reaction, priority journal, screening test, sheep, validation study, Bos, Echinococcus granulosus, Ovis aries, Animals, Camels, Cattle, Cattle Diseases, Echinococcosis, Echinococcus granulosus, Genotype, Iran, Sheep, Sheep Diseases
Subjects: مقالات نمایه شده محققین دانشگاه در سایت ,Web of Science ,Scopus
موارد کلی
Divisions: معاونت تحقیقات و فناوری
Depositing User: GOUMS
Date Deposited: 20 Apr 2015 04:36
Last Modified: 10 May 2015 15:08
URI: http://eprints.goums.ac.ir/id/eprint/1828

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