Open Access
Research (Published online: 22-02-2018)
23. DNA extraction from hydatid cyst protoscolices: Comparison of five different methods
Afshin Barazesh, Bahador Sarkari, Sepideh Ebrahimi and Mehdi Hami
Veterinary World, 11(2): 231-234

Afshin Barazesh: Student Research Committee, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran; The Persian Gulf Tropical Medicine Research Centre, Bushehr University of Medical Sciences, Bushehr, Iran.
Bahador Sarkari: Basic Sciences in Infectious Diseases Research Center, Shiraz University of Medical Sciences, Shiraz, Iran; Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.
Sepideh Ebrahimi: Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.
Mehdi Hami: Technical Deputy of East-Azarbaijan Province, Veterinary Directorate, Iran Veterinary Organization, Iran.

doi: 10.14202/vetworld.2018.231-234

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Article history: Received: 06-11-2017, Accepted: 19-01-2018, Published online: 22-02-2018

Corresponding author: Bahador Sarkari


Citation: Barazesh A, Sarkari B, Ebrahimi S, Hami M (2018) DNA extraction from hydatid cyst protoscolices: Comparison of five different methods, Veterinary World, 11(2): 231-234.

Aim: The current study aimed to find out a simple, practical and high throughput DNA isolation method for extraction of DNA from hydatid cyst samples.

Materials and Methods: Cattle and sheep isolate of hydatid cysts were obtained from the slaughterhouse, and hydatid fluid and protoscolices were collected in a sterile condition. Protoscolices were washed, 3 times with phosphate buffered saline, and DNA was extracted by different methods including manual extraction with freeze/thawing and phenol-chloroform, Triton X-100 extraction, and by a commercial kit (YTA, Yekta Tajhiz Azma, Iran) with three different modifications in the kit's manufacturer instructions. The obtained DNA from the different methods was evaluated by Nanodrop in terms of the yield of DNA and carbohydrates or protein contaminations. To compare the quality of the extracted DNA, two pieces of the mitochondrial genome of Echinococcus granulosus, cox1, and nad1, were polymerase chain reaction (PCR)-amplified, using each of the DNA prepared by different methods. Electrophoresis of PCR products was carried out on the agarose gel.

Results: The DNA extracted by manual method, using phenol/chloroform, had the highest yield, yet with the highest level of protein and carbohydrate contamination. The DNA extracted using two-step incubations, initially at 60°C for 2 h and then overnight at 37°C, was the most purified DNA with the lowest rate of contamination.

Conclusion: Findings of the study demonstrated that modification in the currently available commercially DNA extraction kit resulted in the development of a high throughput DNA isolation method. This method can be recommended for the extraction of DNA from hydatid cysts, especially the cattle isolate where the extraction of DNA in these samples are usually problematic.

Keywords: DNA extraction, hydatid cyst, protoscolices.


1. Deplazes, P., Rinaldi, L., Alvarez, R.C.A., Torgerson, P.R., Harandi, M.F., Romig, T., Antolova, D., Schurer, J.M., Lahmar, S., Cringoli, G., Magambo, J., Thompson, R.C.A. and Jenkins, E.G. (2017) Global distribution of alveolar and cystic echinococcosis. Adv. Parasitol., 95: 315-493. [Crossref] [PubMed]

2. Sarkari, B., Fatemie, S.A., Moshfe, A., Abdolahi, K.S., Savardashtaki, A., Hosseini, F. and Shahbazi, A. (2016) Clinical and molecular evaluation of a case of giant primary splenic hydatid cyst: A case report. Iran J. Parasitol., 11: 585-590. [PubMed] [PMC]

3. Fasihi, H.M., Budke, C.M., Rostami, S. (2012) The monetary burden of cystic echinococcosis in Iran. PLoS Negl. Trop. Dis., 6: e1915. [Crossref] [PubMed] [PMC]

4. Sarkari, B., Sadjjadi, S.M., Beheshtian, M.M., Aghaee, M., and Sedaghat, F. (2010) Human cystic echinococcosis in Yasuj District in Southwest of Iran: An epidemiological study of seroprevalence and surgical cases over a ten-year period. Zoonoses Public Health, 57: 146-150. [Crossref] [PubMed]

5. Siracusano, A., and Bruschi, F. (2006) Cystic echinococcosis: Progress and limits in epidemiology and immunodiagnosis. Parassitologia, 48(1-2): 65-66. [PubMed]

6. Sarkari, B., Hosseini, F., Khabisi, S.A. and Sedaghat, F. (2016) Seroprevalence of cystic echinococcosis in blood donors in Fars province, southern Iran. Parasite Epidemiol. Control, 2: 8-12. [Crossref]

7. Mansouri, M., Sarkari, B. and Mowlavi, G.R. (2016) Helminth parasites of wild boars, sus scrofa, in Bushehr Province, Southwestern Iran. Iran J. Parasitol., 11: 377-382. [PubMed] [PMC]

8. Thompson, R.C. (2017) Biology and systematics of Echinococcus. Adv. Parasitol., 95: 65-109. [Crossref] [PubMed]

9. Sarkari, B., Mansouri, M., Khabisi, S. A., and Mowlavi, G. (2015). Molecular characterization and seroprevalence of Echinococcus granulosus in wild boars (Sus scrofa) in south-western Iran. Ann. Parasitol., 61: 269-273. [PubMed]

10. Yanagida, T., Mohammadzadeh, T., Kamhawi, S., Nakao, M., Sadjjadi, S.M., Hijjawi, N., Abdel-Hafez, S.K., Sako, Y., Okamoto, M. and Ito, A. (2012) Genetic polymorphisms of Echinococcus granulosus sensu stricto in the Middle East. Parasitol. Int., 61: 599-603. [Crossref] [PubMed]

11. Fatemi, E.A., Sarkari, B. and Mikaeili, F. (2018) Genetic variability of antigen B8/1 among Echinococcus granulosus isolates from human, cattle, and sheep in Fars Province, Southern Iran. Rep. Biochem. Mol. Biol., 6: 164-169.

12. Lymbery, A.J. (2017) Phylogenetic pattern, evolutionary processes and species delimitation in the genus Echinococcus. Adv. Parasitol., 95: 111-145. [Crossref] [PubMed]

13. Rahimi, H., Sadjjadi, S. and Sarkari, B. (2011) Performance of antigen B isolated from different hosts and cyst locations in diagnosis of cystic echinococcosis. Iran J. Parasitol., 6: 12-19. [PubMed] [PMC]

14. Sadjjadi, S.M., Sedaghat, F., Hosseini, S.V. and Sarkari, B. (2009) Serum antigen and antibody detection in echinococcosis: Application in serodiagnosis of human hydatidosis. Korean J. Parasitol., 47: 153-157. [Crossref] [PubMed] [PMC]

15. Sarkari, B. and Rezaei, Z. (2015) Immunodiagnosis of human hydatid disease: Where do we stand? World J. Methodol., 5: 185-195. [Crossref] [PubMed] [PMC]

16. Savardashtaki, A., Sarkari, B., Arianfar, F. and Mostafavi-Pour, Z. (2017) Immunodiagnostic value of Echinococcus granulosus recombinant B8/1 subunit of antigen B. Iran J. Immunol., 14: 111-122. [PubMed]

17. Sharbatkhori, M., Kia, E., Harandi, M.F., Jalalizand, N., Zahabiun, F., and Mirhendi, H. (2009) Comparison of five simple methods for DNA extraction from Echinococcus granulosus protoscoleces for PCR-amplification of ribosomal DNA. Iran J. Parasitol., 4: 54-60.

18. Sung, K., Khan, S.A., Nawaz, M.S. and Khan, A.A. (2003) A simple and efficient Triton X-100 boiling and chloroform extraction method of RNA isolation from Gram-positive and Gram-negative bacteria. FEMS Microbiol. Lett., 229: 97-101. [Crossref]