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Genetic variability of E. histolytica strains based on the polymorphism of the SREHP gene using nested PCR-RFLP in Erbil, North Iraq
Corresponding Author(s) : Shler Akram Faqe Mahmood
Cellular and Molecular Biology,
Vol. 66 No. 1: Issue 1
Abstract
E. histolytica is an intestinal parasite that causes asymptomatic infection mostly; however, it may also cause amoebic dysentery and liver abscess. Molecular identification is required in epidemiological studies due to the presence of morphologically identical nonpathogenic species. Therefore, this study was conducted to first evaluate the prevalence rate of E. histolytica among symptomatic individuals of Erbil city, and to investigate the genetic diversity of the parasite in a limited geographic area. Accordingly, a total of 2026 samples were examined microscopically, and confirmed by nested PCR for 18s rRNA gene. The results showed that the prevalence rate of E. histolytica was 1.97% (40 samples) among symptomatic patients. The SREHP gene was used as a marker to show the genetic polymorphism of E. histolytica; however, to compare the genetic diversity of symptomatic with asymptomatic isolates, 57 asymptomatic samples were obtained from our previous study. The amplified products of the SREHP gene were digested by AluI endonuclease, and DNA banding patterns were analysed. Results showed 29 different DNA patterns among the 97 symptomatic and asymptomatic samples, 62 of which shared similar DNA patterns. However, 8 different DNA patterns were observed among asymptomatic samples, whereas 15 distinct patterns were observed among symptomatic isolates. In conclusion, this study found that the prevalence rate of E. histolytica was relatively low; relatively high genetic diversity was observed in a restricted endemic area; with higher rates of variability in symptomatic rather than in asymptomatic isolates, indicating a possible correlation between the genotype of E. histolytica and their clinical outcome.
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- Haque R, Huston CD, Hughes M, Houpt E, Petri WA. Amebiasis. New Engl J Med 2003; 348: 1565-73.
- Paul J, Srivastava S, Bhattacharya S. Molecular methods for diagnosis of entamoeba histolytica in a clinical setting: An overview. Exp Parasitol 2007; 116: 35-43.
- Rahman SMM, Haque R, Roy S, Mondal MMH. Genotyping of entamoeba histolytica by real-time polymerase chain reaction with cyber green i and melting curve analysis. Bangl J Vet Med 2006; 4: 53–60.
- Elbakri A, Samie A, Ezzedine S,Odeh REA. Genetic variability of the serine-rich entamoeba histolytica protein gene in clinical isolates from the united arab emirates. Trop Biomed 2014; 31: 370–37.
- Haghighi A, Kobayashi S, Takeuchi T, Thammapalerd N, Nozaki T. Geographic diversity among genotypes of entamoeba histolytica field isolates. J Clin Microbiol 2003; 41: 3748-56.
- Samie A, Obi CL, Bessong PO, E. Houpt SS, Njayou M, Sabeta C, Mduluza T,Guerrant RL. Entamoeba histolytica: Genetic diversity of African strains based on the polymorphism of the serine-rich protein gene. Exp Parasitol 2008; 118: 354-61.
- Turan H, Karkucak M, Yakut T, Ozsahin M, Gurlevik Z, Yanik ME, Ucgun T, Aliagaoglu C, Yaykasli KO. Does MBL-2 codon 54 polymorphism have a role psoriasis pathogenesis? Int J Dermatol 2014; 53(1):34-38.
- Khairnar K, Parija SC. A novel nested multiplex polymerase chain reaction (PCR) assay for differential detection of entamoeba histolytica, e. Moshkovskii and e. Dispar DNA in stool samples. BMC Microbiol 2007; 7: 47.
- Yaykasli KO, Yaykasli E, Ataoglu S, Ozsahin M, Memisogulları R, í‡elebi E, Uçgun T, Ozcan ME, Uslu M, Yuce H. The frequency of omentin Val109Asp polymorphism and the serum level of omentin in patients with Rheumatoid Arthritis. Acta Medica Mediterranea 2013; 29:521-526.
- Ayeh-Kumi PF, Ali IM, Lockhart LA, Gilchrist CA, Petri WA, Haque R. Entamoeba histolytica: Genetic diversity of clinical isolates from Bangladesh as demonstrated by polymorphisms in the serine-rich gene. Exp Parasitol 2001; 99: 80-8.
- Yaykasli KO, Kayıkçı MA, Yamak N, Soguktas H, Duzenli S, Arslan AO, Metin A, Kaya E, Hatipoglu OF. Polymorphisms in MMP-2 and TIMP-2 in Turkish patients with prostate cancer, Turk J Med Sci 2014; 44:839-843.
- Hussein RA, Al-Mayah QS, Merdaw Ma-Z, Al-Bashier NT, Al-Abbas AA, Jasem IA. Evaluation of multiplex real-time PCR and ELISA in detection of intestinal protozoan parasites from children with diarrheal disease Inter J Adv Res 2015; 3: 782-8.
- Al-Khalidi Kah. Detection of entamoeba histolytica in patients an infected infants with diarrhea in born and children's hospital by classic methods and real-time polymerase chain reaction. J Al Qadisiyah Pure Sci 2016; 21: 27-35.
- Al-Hameedawi JJY. Molecular identification of entamoeba histolytica parasite by using actin and amebapore-a genes. Kufa J for Nursing Sci 2014; 4: 48-55.
- Jaran AS. Prevalence and seasonal variation of human intestinal parasites in patients attending hospital with abdominal symptoms in northern Jordan. E Mediterr Health J 2016; 22: 756-60.
- Nitiema LW, Nordgren J, Ouermi D, Dianou D, Traore AS, Svensson L, Simpore J. Burden of rotavirus and other enteropathogens among children with diarrhea in Burkina Faso. Int J Infect Dis 2011; 15: e646-52.
- Bahrami F, Haghighi A, Zamini G, Khademerfan M. Differential detection of entamoeba histolytica, entamoeba disparandentamoeba moshkovskiiin faecal samples using nested multiplex PCR in west of Iran. Epidemiol Infect 2019; 147.
- Elbakri A, Samie A, Ezzedine S, Odeh RA. Differential detection of entamoeba histolytica, entamoeba dispar and entamoeba moshkovskii in fecal samples by nested PCR in the United Arab Emirates (UAE). Acta Parasitol 2013; 58: 185-90.
- Anuar TS, Al-Mekhlafi HM, Abdul Ghani MK, Azreen SN, Salleh FM, Ghazali N, Bernadus M, Moktar N. Different clinical outcomes of entamoeba histolytica in Malaysia: Does genetic diversity exist?. Korean J Parasitol 2013; 51: 231-6.
- Simonishvili S, Tsanava S, Sanadze K, Chlikadze R, Miskalishvili A, Lomkatsi N, Imnadze P, Petri WA, Jr., Trapaidze N. Entamoeba histolytica: The serine-rich gene polymorphism-based genetic variability of clinical isolates from Georgia. Exp Parasitol 2005; 110: 313-7.
- Tanyüksel M, Ulukanl"¹G"¹L M, Yilmaz H, Güçlü Z, Araz RE, Mert G, Koru í–, William A. Petri J. Genetic variability of the serine-rich gene of entamoeba histolytica in clinical isolates from turkey. Turk J Med Sci 2008; 38: 239-44
- Teixeira JE, Huston CD. Participation of the serine-rich entamoeba histolytica protein in amebic phagocytosis of apoptotic host cells. Infect Immun 2008; 76: 959-66.
- Samuel L.Stanley J, Becker A, Kunz-Jenkins C, Foster L, Li E. Cloning and expression of a membrane antigen of entamoeba histolytica possessing multiple tandem repeats Proc Natl Acad Sci 1990; 87:4976-80.
- Willhoeft U, Tannich E. The electrophoretic karyotype of entamoeba histolytica. Mol Biochem Parasitol 1999; 99: 41-53.
- Rivera WL, Santos SR, Kanbara H. Prevalence and genetic diversity of entamoeba histolytica in an institution for the mentally retarded in the Philippines. Parasitol Res 2006; 98: 106-10.
- Weedall GD, Clark CG, Koldkjaer P, Kay S, Bruchhaus I, Tannich E, Paterson S, Hall N. Genomic diversity of the human intestinal parasite entamoeba histolytica. Genome Biol 2012; 13: R38.
- Weedall GD, Hall N. Evolutionary genomics of entamoeba. Res Microbiol 2011; 162: 637-45.
- Zhang T, Cieslak PR, Stanley SL. Protection of gerbils from amebic liver abscess by immunization with a recombinant entamoeba histolytica antigen. Infect Immun 1994; 62: 1166-70.
- Clark Cg, Diamond Ls. Entamoeba histolytica: A method for isolate identification. Exp Parasitol 1993; 77: 450-5.
References
Haque R, Huston CD, Hughes M, Houpt E, Petri WA. Amebiasis. New Engl J Med 2003; 348: 1565-73.
Paul J, Srivastava S, Bhattacharya S. Molecular methods for diagnosis of entamoeba histolytica in a clinical setting: An overview. Exp Parasitol 2007; 116: 35-43.
Rahman SMM, Haque R, Roy S, Mondal MMH. Genotyping of entamoeba histolytica by real-time polymerase chain reaction with cyber green i and melting curve analysis. Bangl J Vet Med 2006; 4: 53–60.
Elbakri A, Samie A, Ezzedine S,Odeh REA. Genetic variability of the serine-rich entamoeba histolytica protein gene in clinical isolates from the united arab emirates. Trop Biomed 2014; 31: 370–37.
Haghighi A, Kobayashi S, Takeuchi T, Thammapalerd N, Nozaki T. Geographic diversity among genotypes of entamoeba histolytica field isolates. J Clin Microbiol 2003; 41: 3748-56.
Samie A, Obi CL, Bessong PO, E. Houpt SS, Njayou M, Sabeta C, Mduluza T,Guerrant RL. Entamoeba histolytica: Genetic diversity of African strains based on the polymorphism of the serine-rich protein gene. Exp Parasitol 2008; 118: 354-61.
Turan H, Karkucak M, Yakut T, Ozsahin M, Gurlevik Z, Yanik ME, Ucgun T, Aliagaoglu C, Yaykasli KO. Does MBL-2 codon 54 polymorphism have a role psoriasis pathogenesis? Int J Dermatol 2014; 53(1):34-38.
Khairnar K, Parija SC. A novel nested multiplex polymerase chain reaction (PCR) assay for differential detection of entamoeba histolytica, e. Moshkovskii and e. Dispar DNA in stool samples. BMC Microbiol 2007; 7: 47.
Yaykasli KO, Yaykasli E, Ataoglu S, Ozsahin M, Memisogulları R, í‡elebi E, Uçgun T, Ozcan ME, Uslu M, Yuce H. The frequency of omentin Val109Asp polymorphism and the serum level of omentin in patients with Rheumatoid Arthritis. Acta Medica Mediterranea 2013; 29:521-526.
Ayeh-Kumi PF, Ali IM, Lockhart LA, Gilchrist CA, Petri WA, Haque R. Entamoeba histolytica: Genetic diversity of clinical isolates from Bangladesh as demonstrated by polymorphisms in the serine-rich gene. Exp Parasitol 2001; 99: 80-8.
Yaykasli KO, Kayıkçı MA, Yamak N, Soguktas H, Duzenli S, Arslan AO, Metin A, Kaya E, Hatipoglu OF. Polymorphisms in MMP-2 and TIMP-2 in Turkish patients with prostate cancer, Turk J Med Sci 2014; 44:839-843.
Hussein RA, Al-Mayah QS, Merdaw Ma-Z, Al-Bashier NT, Al-Abbas AA, Jasem IA. Evaluation of multiplex real-time PCR and ELISA in detection of intestinal protozoan parasites from children with diarrheal disease Inter J Adv Res 2015; 3: 782-8.
Al-Khalidi Kah. Detection of entamoeba histolytica in patients an infected infants with diarrhea in born and children's hospital by classic methods and real-time polymerase chain reaction. J Al Qadisiyah Pure Sci 2016; 21: 27-35.
Al-Hameedawi JJY. Molecular identification of entamoeba histolytica parasite by using actin and amebapore-a genes. Kufa J for Nursing Sci 2014; 4: 48-55.
Jaran AS. Prevalence and seasonal variation of human intestinal parasites in patients attending hospital with abdominal symptoms in northern Jordan. E Mediterr Health J 2016; 22: 756-60.
Nitiema LW, Nordgren J, Ouermi D, Dianou D, Traore AS, Svensson L, Simpore J. Burden of rotavirus and other enteropathogens among children with diarrhea in Burkina Faso. Int J Infect Dis 2011; 15: e646-52.
Bahrami F, Haghighi A, Zamini G, Khademerfan M. Differential detection of entamoeba histolytica, entamoeba disparandentamoeba moshkovskiiin faecal samples using nested multiplex PCR in west of Iran. Epidemiol Infect 2019; 147.
Elbakri A, Samie A, Ezzedine S, Odeh RA. Differential detection of entamoeba histolytica, entamoeba dispar and entamoeba moshkovskii in fecal samples by nested PCR in the United Arab Emirates (UAE). Acta Parasitol 2013; 58: 185-90.
Anuar TS, Al-Mekhlafi HM, Abdul Ghani MK, Azreen SN, Salleh FM, Ghazali N, Bernadus M, Moktar N. Different clinical outcomes of entamoeba histolytica in Malaysia: Does genetic diversity exist?. Korean J Parasitol 2013; 51: 231-6.
Simonishvili S, Tsanava S, Sanadze K, Chlikadze R, Miskalishvili A, Lomkatsi N, Imnadze P, Petri WA, Jr., Trapaidze N. Entamoeba histolytica: The serine-rich gene polymorphism-based genetic variability of clinical isolates from Georgia. Exp Parasitol 2005; 110: 313-7.
Tanyüksel M, Ulukanl"¹G"¹L M, Yilmaz H, Güçlü Z, Araz RE, Mert G, Koru í–, William A. Petri J. Genetic variability of the serine-rich gene of entamoeba histolytica in clinical isolates from turkey. Turk J Med Sci 2008; 38: 239-44
Teixeira JE, Huston CD. Participation of the serine-rich entamoeba histolytica protein in amebic phagocytosis of apoptotic host cells. Infect Immun 2008; 76: 959-66.
Samuel L.Stanley J, Becker A, Kunz-Jenkins C, Foster L, Li E. Cloning and expression of a membrane antigen of entamoeba histolytica possessing multiple tandem repeats Proc Natl Acad Sci 1990; 87:4976-80.
Willhoeft U, Tannich E. The electrophoretic karyotype of entamoeba histolytica. Mol Biochem Parasitol 1999; 99: 41-53.
Rivera WL, Santos SR, Kanbara H. Prevalence and genetic diversity of entamoeba histolytica in an institution for the mentally retarded in the Philippines. Parasitol Res 2006; 98: 106-10.
Weedall GD, Clark CG, Koldkjaer P, Kay S, Bruchhaus I, Tannich E, Paterson S, Hall N. Genomic diversity of the human intestinal parasite entamoeba histolytica. Genome Biol 2012; 13: R38.
Weedall GD, Hall N. Evolutionary genomics of entamoeba. Res Microbiol 2011; 162: 637-45.
Zhang T, Cieslak PR, Stanley SL. Protection of gerbils from amebic liver abscess by immunization with a recombinant entamoeba histolytica antigen. Infect Immun 1994; 62: 1166-70.
Clark Cg, Diamond Ls. Entamoeba histolytica: A method for isolate identification. Exp Parasitol 1993; 77: 450-5.