Genetic Characteristics of Echinococcus granulosus from Fixed Paraffin-Embedded Tissue Samples in Human Isolates Based on the High-Resolution Melting Point Analysis in Sabzevar, Northeast Iran
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Abstract
Background: There are ten genotypes of Echinococcus granulosus with different intermediate and final hosts affecting the parasite's life cycle and its transmission to humans. Therefore, this study was conducted to determine the genotype of isolated hydatid cysts using the simple and fast high-resolution melting point analysis (HRM) method.
Methods: The paraffin tissue samples of patients who underwent surgery were obtained from the pathology sample bank of Vasei and Emdad Hospitals in Sabzevar, Iran during 2010-2020. The DNA content of the samples was extracted after collecting and determining the characteristics using the DNA extraction kit. PCR was performed on the samples and the presence of the hydatid cyst genome was confirmed using the special Master Kit. Mix PCR of Solis Biodyne Company and Real-Time device (Bio-Rad) were used, and the genetic identity of hydatid cysts were determined.
Results: Out of 33 paraffin samples, 21 samples contained hydatid cyst DNA, two of which were from the brain and 19 from the liver tissues; 12 samples did not contain hydatid cyst DNAs. All liver samples were from sheep species (G1), and the brain samples were from buffalo species (G3). Therefore, 9.53% of the Echinococcus species collected were buffalo (G3), and 90.47% were sheep (G1) strain.
Conclusion: Based on previous patterns, HRM methods can be used for easy and quick identification of Echinococcus strains. The G1 strain was the dominant strain causing hydatid cyst in different human organs, including the liver and brain.
1. Eckert J, Deplazes P. Biological, epidemiological, and clinical aspects of echinococcosis, a zoonosis of increasing concern. Clin Microbiol Rev. 2004;17(1):107-35.
2. Moro P, Schantz PM. Echinococcosis: a review. Int J Infect Dis. 2009;13(2):125-33.
3. Murray CJ, Vos T, Lozano R, et al. Disability-adjusted life years (DALYs) for 291 diseases and injuries in 21 regions, 1990–2010: a systematic analysis for the Global Burden of Disease Study 2010. Lancet. 2012;380(9859):2197-223.
4. Thompson RC. Biology and Systematics of Echinococcus. Adv Parasitol. 2017;95:65-109.
5. Haniloo A, Farhadi M, Fazaeli A, Nouri, et al. Genotype characterization of hydatid cysts isolated from Zanjan using PCR-RFLP technique. J Adv Med Biomed Res. 2013;21(84):57–65.
6. Abbasi I, Branzburg A, Campos-Ponce M, et al. Copro-diagnosis of Echinococcus granulosus infection in dogs by amplification of a newly identified repeated DNA sequence. Am J Trop Med Hyg. 2003;69(3):324-30.
7. Nakao M, McManus DP, Schantz PM, et al. A molecular phylogeny of the genus Echinococcus inferred from complete mitochondrial genomes. Parasitology. 2007;134(Pt 5):713-22.
8. Zamand S, Alidadi H, Najafpoor A, Taghavimanesh V, Akbari Sharak N, Najafi Saleh H. Data on trend changes of drinking groundwater resources quality in Sabzevar city (A case study). Data Brief. 2018; 20: 889-893.
9. Rostami S, Shariat Torbaghan S, Dabiri S, et al. Genetic characterization of Echinococcus granulosus from a large number of formalin-fixed, paraffin-embedded tissue samples of human isolates in Iran. Am J Trop Med Hyg. 2015;92(3):588-94.
10. Metwally DM, Qassim LE, Al-Turaiki IM, et al. Gene-based molecular analysis of COX1 in Echinococcus granulosus cysts isolated from naturally infected livestock in Riyadh, Saudi Arabia. PLoS One. 2018;13(4):e0195016.
11. Borji H, Bahar MM, Naghibi A, et al. Genotyping of human cystic echinococcosis in northeastern Iran. Ann Parasitol. 2018;64(4):323-329.
12. Dalimi A, Shamsi M, Khosravi A,et al. Genotyping Echinococcus granulosus from Canine Isolates in Ilam Province, West of Iran. Iran J Parasitol. 2017;12(4):614-621.
13. Abedi B, Maghsood AH, Khansarinejad B, et al. Genotyping of Echinococcus granulosus isolates from livestock based on mitochondrial cox1 gene, in the Markazi province, Iran. J Parasit Dis. 2019;43(4):592-596.
14. Nikmanesh B, Mirhendi H, Ghalavand Z, et al. Genotyping of Echinococcus granulosus Isolates from Human Clinical Samples Based on Sequencing of Mitochondrial Genes in Iran, Tehran. Iran J Parasitol. 2014;9(1):20-7.
15. Berenji F, Shamsian SA, Nouri Daloee M, et al. Genotyping of Echinococcus granulosus Isolates from Human in Khorasan Province, North-Eastern Iran. Iran J Parasitol. 2019;14(1):52-58.
16. Pezeshki A, Akhlaghi L, Sharbatkhori M, et al. Genotyping of Echinococcus granulosus from domestic animals and humans from Ardabil Province, northwest Iran. J Helminthol. 2013;87(4):387-91.
17. Babaei Z, Taherkhani R, Barazesh A, et al. Genotyping and phylogenetic analysis of hydatid cysts isolated from livestock in Bushehr province, Iran. J Parasit Dis. 2021;45(1):197-203.
18. Zarei, Z., Ghalehbin, B.M., Akhoundi, B. et al. Genotyping of Echinococcus granulosus isolated from canine in Northwest Iran. J Parasit Dis. 2023; 47(4):757–761.
19. Shahbazi AE, Saidijam M, Maghsood AH, et al. Genotyping of Fresh and Paraffinized Human Hydatid Cysts Using nad1 and cox1 Genes in Hamadan Province, West of Iran. Iran J Parasitol. 2020;15(2):259-265.
20. Shahabi S, Sarkari B, Barazesh A. Echinococcus granulosus sensu stricto G1 is the predominant genotype in human and livestock isolates from Turkey and Iran, based on mitochondrial nad5 gene differentiation. Parasit Vectors. 2021;14(1):369.
21. Salehi M, Yaghfoori S, Bahari P, et al. Molecular Characterization of Echinococcus granulosus Sensu Lato from Livestock in North Khorasan Province, Iran. Iran J Parasitol. 2018;13(4):577-586.
22. Zarrabi Ahrabi S, Madani R, Shemshadi B, et al. Genetic Affinity of Echinococcus granulosus protoscolex in Human and Sheep in East Azerbaijan, Iran. Arch Razi Inst. 2020;75(1):47-54.
23. Gholami S, Jahandar H, Abastabar M, et al. Echinococcus granulosus Sensu Stricto in Dogs and Jackals from Caspian Sea Region, Northern Iran. Iran J Parasitol. 2016;11(2):186-194.
24. Ghabdian S, Borji H, Naghibi A. Molecular identification of Echinococcus granulosus strain in stray dogs from Northeastern Iran. Vet Parasitol Reg Stud Reports. 2017; 9:6-8.
25. Kargar S, Dalimi A, Eslami G, et al. Cystic Echinococcosis in Central Iran: G1 and G6 Genotypes in Patients. Surg Infect (Larchmt). 2022; 23(5):451-457.
26. Karamian M, Haghighi F, Hemmati M. Genotyping of Cystic Hydatidosis Agents in Birjand, Eastern Iran. Mod Care J. 2017;14(3):e64531.
27. Gorgani-Firouzjaee T, Kalantrai N, Ghaffari S, et al. Genotype characterization of livestock and human cystic echinococcosis in Mazandaran province, Iran. J Helminthol. 2019;93(2):255-259.
28. Sharbatkhori M, Mirhendi H, Jex AR, et al. Genetic categorization of Echinococcus granulosus from humans and herbivorous hosts in Iran using an integrated mutation scanning-phylogenetic approach. Electrophoresis. 2009;30(15):2648-55.
29. Zhang L, Eslami A, Hosseini SH, et al. Indication of the presence of two distinct strains of Echinococcus granulosus in Iran by mitochondrial DNA markers. Am J Trop Med Hyg. 1998;59(1):171-4.
30. Vahedi A, Mahdavi M, Ghazanchaei A, et al. Genotypic characteristics of hydatid cysts isolated from humans in East Azerbaijan Province (2011-2013). J Anal Res Clin Med. 2014; 2(3):152-157.
31. Nikmanesh B, Mirhendi H, Ghalavand Z, et al. Genotyping of Echinococcus granulosus Isolates from Human Clinical Samples Based on Sequencing of Mitochondrial Genes in Iran, Tehran. Iran J Parasitol. 2014;9(1):20-7.
32. Gholami Sh, Sosari M, Fakhar M, et al. Molecular Characterization of Echinococcus granulosus from Hydatid Cysts Isolated from Human and Animals in Golestan Province, North of Iran. Iran J Parasitol. 2012;7(4):8-16.
33. Pezeshki A, Akhlaghi L, Sharbatkhori M, et al. Genotyping of Echinococcus granulosus from domestic animals and humans from Ardabil Province, northwest Iran. J Helminthol. 2013;87(4):387-91.
34. Sadjjadi SM, Mikaeili F, Karamian M, et al. Evidence that the Echinococcus granulosus G6 genotype has an affinity for the brain in humans. Int J Parasitol. 2013;43(11):875-7.
35. Hajialilo E, Harandi MF, Sharbatkhori M, et al. Genetic characterization of Echinococcus granulosus in camels, cattle and sheep from the south-east of Iran indicates the presence of the G3 genotype. J Helminthol. 2012;86(3):263-70.
36. Elyasi H, Golmohammadi R. Genetic Identification of Echinococcus granulosus Isolates from Domestic Animals in Sabzevar, Northeast Iran. Iran J Parasitol. 2020;15(4):621-623.
37. Sadri A, Moshfe A, Doosti A, et al. Characterization of Isolated Hydatid Cyst from Slaughtered Livestock in Yasouj Industrial Slaughterhouse by PCR-RFLP. Armaghan Danesh, 2012; 17(3): 243-252.
38. Ergin S, Saribas S, Yuksel P, et al. Genotypic characterization of Echinococcus granulosus isolated from human in Turkey. Afr J Microbiol Res. 2010; 4(7):551–555
39. Khademvatan S, Yousefi E, Rafiei A, et al. Molecular characterization of livestock and human isolates of Echinococcus granulosus from south-west Iran. J Helminthol. 2013;87(2):240-4.
40. Macin S, Orsten S, Samadzade R, et al. Human and animal cystic echinococcosis in Konya, Turkey: molecular identification and the first report of E. equinus from human host in Turkey. Parasitol Res. 2021;120(2):563-568.
41. Shafiei R, Ghatee MA, Jafarzadeh F, et al. Genotyping and phylogenetic analysis of unusually located hydatid cysts isolated from humans in north-east Iran. J Helminthol. 2019;94:e64.
42. Hedayati Z, Daryani A, Sarvi S, et al. Molecular Genotyping of the Human Cystic Echinococcosis in Mazandaran Province, North of Iran. Iran J Parasitol. 2019;14(1):151-158.
2. Moro P, Schantz PM. Echinococcosis: a review. Int J Infect Dis. 2009;13(2):125-33.
3. Murray CJ, Vos T, Lozano R, et al. Disability-adjusted life years (DALYs) for 291 diseases and injuries in 21 regions, 1990–2010: a systematic analysis for the Global Burden of Disease Study 2010. Lancet. 2012;380(9859):2197-223.
4. Thompson RC. Biology and Systematics of Echinococcus. Adv Parasitol. 2017;95:65-109.
5. Haniloo A, Farhadi M, Fazaeli A, Nouri, et al. Genotype characterization of hydatid cysts isolated from Zanjan using PCR-RFLP technique. J Adv Med Biomed Res. 2013;21(84):57–65.
6. Abbasi I, Branzburg A, Campos-Ponce M, et al. Copro-diagnosis of Echinococcus granulosus infection in dogs by amplification of a newly identified repeated DNA sequence. Am J Trop Med Hyg. 2003;69(3):324-30.
7. Nakao M, McManus DP, Schantz PM, et al. A molecular phylogeny of the genus Echinococcus inferred from complete mitochondrial genomes. Parasitology. 2007;134(Pt 5):713-22.
8. Zamand S, Alidadi H, Najafpoor A, Taghavimanesh V, Akbari Sharak N, Najafi Saleh H. Data on trend changes of drinking groundwater resources quality in Sabzevar city (A case study). Data Brief. 2018; 20: 889-893.
9. Rostami S, Shariat Torbaghan S, Dabiri S, et al. Genetic characterization of Echinococcus granulosus from a large number of formalin-fixed, paraffin-embedded tissue samples of human isolates in Iran. Am J Trop Med Hyg. 2015;92(3):588-94.
10. Metwally DM, Qassim LE, Al-Turaiki IM, et al. Gene-based molecular analysis of COX1 in Echinococcus granulosus cysts isolated from naturally infected livestock in Riyadh, Saudi Arabia. PLoS One. 2018;13(4):e0195016.
11. Borji H, Bahar MM, Naghibi A, et al. Genotyping of human cystic echinococcosis in northeastern Iran. Ann Parasitol. 2018;64(4):323-329.
12. Dalimi A, Shamsi M, Khosravi A,et al. Genotyping Echinococcus granulosus from Canine Isolates in Ilam Province, West of Iran. Iran J Parasitol. 2017;12(4):614-621.
13. Abedi B, Maghsood AH, Khansarinejad B, et al. Genotyping of Echinococcus granulosus isolates from livestock based on mitochondrial cox1 gene, in the Markazi province, Iran. J Parasit Dis. 2019;43(4):592-596.
14. Nikmanesh B, Mirhendi H, Ghalavand Z, et al. Genotyping of Echinococcus granulosus Isolates from Human Clinical Samples Based on Sequencing of Mitochondrial Genes in Iran, Tehran. Iran J Parasitol. 2014;9(1):20-7.
15. Berenji F, Shamsian SA, Nouri Daloee M, et al. Genotyping of Echinococcus granulosus Isolates from Human in Khorasan Province, North-Eastern Iran. Iran J Parasitol. 2019;14(1):52-58.
16. Pezeshki A, Akhlaghi L, Sharbatkhori M, et al. Genotyping of Echinococcus granulosus from domestic animals and humans from Ardabil Province, northwest Iran. J Helminthol. 2013;87(4):387-91.
17. Babaei Z, Taherkhani R, Barazesh A, et al. Genotyping and phylogenetic analysis of hydatid cysts isolated from livestock in Bushehr province, Iran. J Parasit Dis. 2021;45(1):197-203.
18. Zarei, Z., Ghalehbin, B.M., Akhoundi, B. et al. Genotyping of Echinococcus granulosus isolated from canine in Northwest Iran. J Parasit Dis. 2023; 47(4):757–761.
19. Shahbazi AE, Saidijam M, Maghsood AH, et al. Genotyping of Fresh and Paraffinized Human Hydatid Cysts Using nad1 and cox1 Genes in Hamadan Province, West of Iran. Iran J Parasitol. 2020;15(2):259-265.
20. Shahabi S, Sarkari B, Barazesh A. Echinococcus granulosus sensu stricto G1 is the predominant genotype in human and livestock isolates from Turkey and Iran, based on mitochondrial nad5 gene differentiation. Parasit Vectors. 2021;14(1):369.
21. Salehi M, Yaghfoori S, Bahari P, et al. Molecular Characterization of Echinococcus granulosus Sensu Lato from Livestock in North Khorasan Province, Iran. Iran J Parasitol. 2018;13(4):577-586.
22. Zarrabi Ahrabi S, Madani R, Shemshadi B, et al. Genetic Affinity of Echinococcus granulosus protoscolex in Human and Sheep in East Azerbaijan, Iran. Arch Razi Inst. 2020;75(1):47-54.
23. Gholami S, Jahandar H, Abastabar M, et al. Echinococcus granulosus Sensu Stricto in Dogs and Jackals from Caspian Sea Region, Northern Iran. Iran J Parasitol. 2016;11(2):186-194.
24. Ghabdian S, Borji H, Naghibi A. Molecular identification of Echinococcus granulosus strain in stray dogs from Northeastern Iran. Vet Parasitol Reg Stud Reports. 2017; 9:6-8.
25. Kargar S, Dalimi A, Eslami G, et al. Cystic Echinococcosis in Central Iran: G1 and G6 Genotypes in Patients. Surg Infect (Larchmt). 2022; 23(5):451-457.
26. Karamian M, Haghighi F, Hemmati M. Genotyping of Cystic Hydatidosis Agents in Birjand, Eastern Iran. Mod Care J. 2017;14(3):e64531.
27. Gorgani-Firouzjaee T, Kalantrai N, Ghaffari S, et al. Genotype characterization of livestock and human cystic echinococcosis in Mazandaran province, Iran. J Helminthol. 2019;93(2):255-259.
28. Sharbatkhori M, Mirhendi H, Jex AR, et al. Genetic categorization of Echinococcus granulosus from humans and herbivorous hosts in Iran using an integrated mutation scanning-phylogenetic approach. Electrophoresis. 2009;30(15):2648-55.
29. Zhang L, Eslami A, Hosseini SH, et al. Indication of the presence of two distinct strains of Echinococcus granulosus in Iran by mitochondrial DNA markers. Am J Trop Med Hyg. 1998;59(1):171-4.
30. Vahedi A, Mahdavi M, Ghazanchaei A, et al. Genotypic characteristics of hydatid cysts isolated from humans in East Azerbaijan Province (2011-2013). J Anal Res Clin Med. 2014; 2(3):152-157.
31. Nikmanesh B, Mirhendi H, Ghalavand Z, et al. Genotyping of Echinococcus granulosus Isolates from Human Clinical Samples Based on Sequencing of Mitochondrial Genes in Iran, Tehran. Iran J Parasitol. 2014;9(1):20-7.
32. Gholami Sh, Sosari M, Fakhar M, et al. Molecular Characterization of Echinococcus granulosus from Hydatid Cysts Isolated from Human and Animals in Golestan Province, North of Iran. Iran J Parasitol. 2012;7(4):8-16.
33. Pezeshki A, Akhlaghi L, Sharbatkhori M, et al. Genotyping of Echinococcus granulosus from domestic animals and humans from Ardabil Province, northwest Iran. J Helminthol. 2013;87(4):387-91.
34. Sadjjadi SM, Mikaeili F, Karamian M, et al. Evidence that the Echinococcus granulosus G6 genotype has an affinity for the brain in humans. Int J Parasitol. 2013;43(11):875-7.
35. Hajialilo E, Harandi MF, Sharbatkhori M, et al. Genetic characterization of Echinococcus granulosus in camels, cattle and sheep from the south-east of Iran indicates the presence of the G3 genotype. J Helminthol. 2012;86(3):263-70.
36. Elyasi H, Golmohammadi R. Genetic Identification of Echinococcus granulosus Isolates from Domestic Animals in Sabzevar, Northeast Iran. Iran J Parasitol. 2020;15(4):621-623.
37. Sadri A, Moshfe A, Doosti A, et al. Characterization of Isolated Hydatid Cyst from Slaughtered Livestock in Yasouj Industrial Slaughterhouse by PCR-RFLP. Armaghan Danesh, 2012; 17(3): 243-252.
38. Ergin S, Saribas S, Yuksel P, et al. Genotypic characterization of Echinococcus granulosus isolated from human in Turkey. Afr J Microbiol Res. 2010; 4(7):551–555
39. Khademvatan S, Yousefi E, Rafiei A, et al. Molecular characterization of livestock and human isolates of Echinococcus granulosus from south-west Iran. J Helminthol. 2013;87(2):240-4.
40. Macin S, Orsten S, Samadzade R, et al. Human and animal cystic echinococcosis in Konya, Turkey: molecular identification and the first report of E. equinus from human host in Turkey. Parasitol Res. 2021;120(2):563-568.
41. Shafiei R, Ghatee MA, Jafarzadeh F, et al. Genotyping and phylogenetic analysis of unusually located hydatid cysts isolated from humans in north-east Iran. J Helminthol. 2019;94:e64.
42. Hedayati Z, Daryani A, Sarvi S, et al. Molecular Genotyping of the Human Cystic Echinococcosis in Mazandaran Province, North of Iran. Iran J Parasitol. 2019;14(1):151-158.
| Files | ||
| Issue | Vol 19 No 3 (2024) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijpa.v19i3.16388 | |
| Keywords | ||
| Echinococcus granulosus Genotyping Hydatid cyst High-resolution melting point analysis | ||
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How to Cite
1.
Elyasi H, Farbodnia T, Javaheri E. Genetic Characteristics of Echinococcus granulosus from Fixed Paraffin-Embedded Tissue Samples in Human Isolates Based on the High-Resolution Melting Point Analysis in Sabzevar, Northeast Iran. Iran J Parasitol. 2024;19(3):305-313.
