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Evaluation of Three PCR Assays for the Identification of the Sheep Strain (Genotype 1) of Echinococcus granulosus in Canid Feces and Parasite Tissues

The performance of 3 PCR assays for the identification of the G1 sheep genotype of Echinococcus granulosus was evaluated using tissue and canid fecal samples. The "Dinkel" and "tefani" primers were the most sensitive in detecting E. granulosus DNA in feces of necropsied dogs (73.7% and 100%, respectively). The "Abbasi" primers detected 52.6% of E. granulosus infected dogs but were the most species-specific, cross-reacting only with Echinococcus shiquicus (tissue 90.9%; feces 75%). The tefani primers were the least specific (tissue, 27.3%; feces, 25%) for E. granulosus. The Dinkel primers also showed inter-species cross-reactivity (tissue, 63.6%; feces, 100%) but were found to be strain-specific for the E. granulosus G1 sheep genotype. Improvement of PCR tests for Echinococcus species and subspecific variants should rely on the use of less-conserved genes and development of protocols that improve the quality and quantity of DNA extracted from feces.

Received November 13, 2007. Accepted for publication February 8, 2008.

Acknowledgments: The laboratory technical support of Mrs. Helen Bradshaw is gratefully acknowledged. Material used in this study was kindly provided by Kevin Shaddick (protoscoleces from horse hydatid G4, Bristol abattoir, Bristol, U.K.), A. Dinkel (E. granulosus G5 DNA, G6 and G7 protoscoleces), B. Gottstein (E. granulosus G5 DNA), P. Dubinsky (E. granulosus G7, protoscoleces and germinal layer), D. McManus (E. granulosus G8 protoscoleces), A. Lavikainen and A. Oksanen (E. granulosus G10 protoscoleces), S. Abdel-Hafez (for access to a dog necropsy isolated from Jordan), Paul Torgerson (feces from dogs experimentally infected with T. multiceps and adult E. granulosus from Kazakhstan), and H. Garcia (T. solium cysts and adult worms, Peru).

Financial support: The research described was supported by a grant (RO1 TW001565) from the Fogarty International Centre.

The content is solely the responsibility of the authors and does not necessarily represent the official views of the Fogarty International Centre or the National Institutes of Health (Principal Investigator, Philip S. Craig).

^* Address correspondence to Belgees S. Boufana, Biomedical Sciences Research Institute, University of Salford, United Kingdom. E-mail: b.boufana{at}salford.ac.uk

Authors’ addresses: Belgees S. Boufana and Philip S. Craig, Cestode Zoonoses Research Group, Biomedical Sciences Research Institute, University of Salford, United Kingdom, Tel: 44-0161-295 4299, Fax: 44-0161-295 5129, E-mail: B.Boufana{at}salford.ac.uk. Maiza Campos-Ponce, Department of Infectious Diseases, Institute for Health Sciences, Vrije Universiteit Amsterdam, De Boelelaan 1085, 1081 HV Amsterdam, The Netherlands. Ariel Naidich, Departamento de Parasitología, Instituto Nacional de Enfermedades Infecciosas, Av. Velez Sarsfield 563, CP 1281, Capital Federal, Argentina. Imad Buishi, El-Fateh University, PO Box 606, Tripoli, Libya. Selma Lahmar, Service de Parasitologie, Ecole Nationale de Médecine Vétérinaire, 2020 Sidi Thabet, Tunisia. Eberhard Zeyhle, Hydatid Unit, African Medical and Research Foundation, PO Box 30125, Nairobi, Kenya. David J. Jenkins, Australian Hydatid Control and Epidemiology Program, 12 Mildura Street, Fyshwick, ACT 2609, Australia. Benoit Combes, Entente Interdepartementale de Lutte Contre la Rage et Autres Zoonoses, Domaine de Pixérécourt, 54220 Malzeville, France. Hao Wen, First Teaching Hospital, Xinjiang Medical University, Urumqi, Xinjiang, China. Ning Xiao, Minoru Nakao, and Akira Ito, Department of Parasitology, Asahikawa Medical College, Asahikawa 078-8510, Japan. Jiamin Qiu, Institute of Parasitic Diseases, Sichuan Centre for Disease Control and Prevention, Chengdu, Sichuan 610041 China.

Reprint requests: Belgees S. Boufana, Biomedical Sciences Research Institute, University of Salford, United Kingdom, Tel: 44-0161-295 4299, Fax: 44-0161-295 5129, E-mail: B.Boufana{at}salford.ac.uk.