Specific Detection of Fasciola hepatica and F. gigantica in Infected Domesticated Animals Using High-Resolution Melting Analysis (HRM)
-
Zeinab MOGHADAMIZAD
,
Ahmad HOSSEINI-SAFA
,
Mehdi MOHEBALI
,
Peyman HEYDARIAN
,
Mojgan ARYAEIPOUR
,
Mohammad Bagher ROKNI
Abstract
Background: It is difficult to make an exact morphological distinction between Fasciola hepatica and Fasciola gigantica. We used High Resolution Melting analysis (HRM) method to differentiate the F. hepatica species from F. gigantica in order to differentiate them.
Methods: Overall, 80 adult liver flukes were collected from infected slaughtered animals including cattle, sheep and goats from Lorestan Province, western Iran from Sep 2015 to Aug 2017. Genomic DNA was extracted using commercial DNA extraction kit. The multilocus sequences of mDNA including COX1, COX3 and ND6 were amplified employing real-time PCR & HRM analysis. Specific and universal primer pairs were designed for differentiation Fasciola spp.
Results: Universal primers cannot be used to distinguish between these two species, but in the contrary, specific primer pairs of each species could differentiate them properly. Molecular identification using specific primer pairs were consistent.
Conclusion: HRM is a simple, fast and reliable method for detecting and differentiating F. hepatica from F. gigantica and can be used for diagnostic and epidemiological purposes.
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13. MAS CS, Bargues M (1997). Human liver flukes: a review. Res Rev Parasitol, 57(3-4):145–218.
14. Ashrafi K (2004). A survey on human and animal fascioliasis and genotypic and phenotypic characteristics of fasciolids and their relationship with lymnaeid snails in Gilan province, northern Iran [Ph. D thesis]. School of Public Health, Tehran University of Medical Sciences, Iran.
15. Esteban JG, Bargues MD, Mas-Coma S (1998). Geographical distribution, diagnosis and treatment of human fascioliasis: a review. Res Rev Parasitol, 58 (I): 13-42.
16. Marcilla A, Bargues M, Mas-Coma S (2002). A PCR-RFLP assay for the distinction between Fasciola hepatica and Fasciola gigantica. Mol Cell Probes, 16(5):327-333.
17. Hosseini-Safa A, Rokni MB, Mosawi SH, Heydarian P, Azizi H, Davari A, Aryaiepour M (2019). High-Resolution Melting Analysis as an Appropriate Method to Differentiate between Fasciola hepatica and F. gigantica. Iran J Public Health, 48(3):501-507.
18. Rokni MB, Mirhendi H, Mizani A, Mohebali M, Sharbatkhori M, Kia EB, Abdoli H, Izadi S (2010). Identification and differentiation of Fasciola hepatica and Fasciola gigantica using a simple PCR-restriction enzyme method. Exp Parasitol, 124(2):209-213.
19. Saki J, Khademvatan S, Yousefi E (2011). Molecular identification of animal Fasciola isolates in southwest of Iran. Australian Journal of Basic and Applied Sciences, 5(11):1878-1883.
20. Hosseini-Safa A, Mohebali M, Hajjaran H, Akhoundi B, Zarei Z, Arzamani K, Davari A (2018). High resolution melting analysis as an accurate method for identifying Leishmania infantum in canine serum samples. J Vector Borne Dis, 55(4):315-320.
21. Pestechian N, Safa AH, Tajedini M, et al (2014). Genetic diversity of Echinococcus granulosus in center of Iran. Korean J Parasitol, 52(4):413-18.
22. Omar MA, Metwally AM, Sultan K (2013). Molecular and phylogenetic status of Fasciola sp., of cattle in Qena, Upper Egypt. Pak J Biol Sci, PJBS, 16(15):726-730.
23. Shoriki T, Ichikawa-Seki M, Devkota B, Rana HB, Devkota SP, Humagain SK, Itagaki T (2014). Molecular phylogenetic identification of Fasciola flukes in Nepal. Parasitol Int, 63(6):758-762.
24. Shafiei R, Sarkari B, Sadjjadi SM, Mowlavi GR, Moshfe A (2014). Molecular and morphological characterization of Fasciola spp. isolated from different host species in a newly emerging focus of human fascioliasis in Iran. Vet Med Int, 2014:405740.
25. Bozorgomid A, Nazari N, Rahimi H, et al (2016). Molecular characterization of animal Fasciola spp. isolates from Kermanshah, Western Iran. Iran J Public Health, 45(10):1315-21.
2. Mahami-Oskouei M, Dalimi A, Forouzandeh-Moghadam M, Rokni M (2011). Molecular identification and differentiation of Fasciola isolates using PCR-RFLP method based on internal transcribed spacer (ITS1, 5.8 S rDNA, ITS2). Iran J Parasitol, 6(3):35-42.
3. Rim H-J, Farag H, Sornmani S, Cross J (1994). Food-borne trematodes: ignored or emerging? Elsevier Current Trends.
4. Valero MA, Bargues MD, Khoubbane M, et al (2016). Higher physiopathogenicity by Fasciola gigantica than by the genetically close F. hepatica: experimental long-term follow-up of biochemical markers. Trans R Soc Trop Med Hyg, 110 (1):55-66.
5. Amor N, Halajian A, Farjallah S, Merella P, Said K, Slimane BB (2011). Molecular characterization of Fasciola spp. from the endemic area of northern Iran based on nuclear ribosomal DNA sequences. Exp Parasitol, 128(3):196-204.
6. Peng M, Ichinomiya M, Ohtori M, Ichikawa M, Shibahara T, Itagaki T (2009). Molecular characterization of Fasciola hepatica, Fasciola gigantica, and aspermic Fasciola sp. in China based on nuclear and mitochondrial DNA. Parasitol Res, 105(3):809-15.
7. Agatsuma T, Arakawa Y, Iwagami M, Honzako Y, Cahyaningsih U, Kang S-Y, Hong S-J (2000). Molecular evidence of natural hybridization between Fasciola hepatica and F. gigantica. Parasitol Int, 49(3):231-238.
8. Itagaki T, Kikawa M, Sakaguchi K, Shimo J, Terasaki K, Shibahara T, Fukuda K (2005). Genetic characterization of parthenogenic Fasciola sp. in Japan on the basis of the sequences of ribosomal and mitochondrial DNA. Parasitology, 131:679-685.
9. Itagaki T, Sakaguchi K, Terasaki K, Sasaki O, Yoshihara S, Van Dung T (2009). Occurrence of spermic diploid and aspermic triploid forms of Fasciola in Vietnam and their molecular characterization based on nuclear and mitochondrial DNA. Parasitol Int, 58(1):81-85.
10. Le TH, Van De N, Agatsuma T, Nguyen TGT, Nguyen QD, McManus DP, Blair D (2008). Human fascioliasis and the presence of hybrid/introgressed forms of Fasciola hepatica and Fasciola gigantica in Vietnam. Int J Parasitol., 38(6):725-730.
11. Amer S, Dar Y, Ichikawa M, Fukuda Y, Tada C, Itagaki T, Nakai Y (2011). Identification of Fasciola species isolated from Egypt based on sequence analysis of genomic (ITS1 and ITS2) and mitochondrial (NDI and COI) gene markers. Parasitol Int, 60(1):5-12.
12. Periago M, Valero M, El Sayed M, et al (2008). First phenotypic description of Fasciola hepatica/ Fasciola gigantica intermediate forms from the human endemic area of the Nile Delta, Egypt. Infect Genet Evol, 8(1):51-58.
13. MAS CS, Bargues M (1997). Human liver flukes: a review. Res Rev Parasitol, 57(3-4):145–218.
14. Ashrafi K (2004). A survey on human and animal fascioliasis and genotypic and phenotypic characteristics of fasciolids and their relationship with lymnaeid snails in Gilan province, northern Iran [Ph. D thesis]. School of Public Health, Tehran University of Medical Sciences, Iran.
15. Esteban JG, Bargues MD, Mas-Coma S (1998). Geographical distribution, diagnosis and treatment of human fascioliasis: a review. Res Rev Parasitol, 58 (I): 13-42.
16. Marcilla A, Bargues M, Mas-Coma S (2002). A PCR-RFLP assay for the distinction between Fasciola hepatica and Fasciola gigantica. Mol Cell Probes, 16(5):327-333.
17. Hosseini-Safa A, Rokni MB, Mosawi SH, Heydarian P, Azizi H, Davari A, Aryaiepour M (2019). High-Resolution Melting Analysis as an Appropriate Method to Differentiate between Fasciola hepatica and F. gigantica. Iran J Public Health, 48(3):501-507.
18. Rokni MB, Mirhendi H, Mizani A, Mohebali M, Sharbatkhori M, Kia EB, Abdoli H, Izadi S (2010). Identification and differentiation of Fasciola hepatica and Fasciola gigantica using a simple PCR-restriction enzyme method. Exp Parasitol, 124(2):209-213.
19. Saki J, Khademvatan S, Yousefi E (2011). Molecular identification of animal Fasciola isolates in southwest of Iran. Australian Journal of Basic and Applied Sciences, 5(11):1878-1883.
20. Hosseini-Safa A, Mohebali M, Hajjaran H, Akhoundi B, Zarei Z, Arzamani K, Davari A (2018). High resolution melting analysis as an accurate method for identifying Leishmania infantum in canine serum samples. J Vector Borne Dis, 55(4):315-320.
21. Pestechian N, Safa AH, Tajedini M, et al (2014). Genetic diversity of Echinococcus granulosus in center of Iran. Korean J Parasitol, 52(4):413-18.
22. Omar MA, Metwally AM, Sultan K (2013). Molecular and phylogenetic status of Fasciola sp., of cattle in Qena, Upper Egypt. Pak J Biol Sci, PJBS, 16(15):726-730.
23. Shoriki T, Ichikawa-Seki M, Devkota B, Rana HB, Devkota SP, Humagain SK, Itagaki T (2014). Molecular phylogenetic identification of Fasciola flukes in Nepal. Parasitol Int, 63(6):758-762.
24. Shafiei R, Sarkari B, Sadjjadi SM, Mowlavi GR, Moshfe A (2014). Molecular and morphological characterization of Fasciola spp. isolated from different host species in a newly emerging focus of human fascioliasis in Iran. Vet Med Int, 2014:405740.
25. Bozorgomid A, Nazari N, Rahimi H, et al (2016). Molecular characterization of animal Fasciola spp. isolates from Kermanshah, Western Iran. Iran J Public Health, 45(10):1315-21.
| Files | ||
| Issue | Vol 49 No 3 (2020) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijph.v49i3.3149 | |
| Keywords | ||
| Fasciola hepatica Fasciola gigantica Fasciolosis Polymerase chain reaction (PCR) | ||
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How to Cite
1.
MOGHADAMIZAD Z, HOSSEINI-SAFA A, MOHEBALI M, HEYDARIAN P, ARYAEIPOUR M, ROKNI MB. Specific Detection of Fasciola hepatica and F. gigantica in Infected Domesticated Animals Using High-Resolution Melting Analysis (HRM). Iran J Public Health. 2020;49(3):521-529.
