Review Article

The Role of Molecular Techniques on Malaria Control and Elimination Programs in Iran: A Review Article


Background: The aim of this review was to describe the application of molecular methods in epidemiological aspects of malaria vectors, parasites, and human hosts in Iran and their critical role in malaria control and elimination programs.

Methods: Medline, EMBASE, Web of Science, Scopus, and Google Scholar databases were searched systematically for original published papers on PCR, the molecular identification of malaria vectors, the molecular epidemiology of malaria, insecticide resistance, and drug-resistant parasites, in Iran. In total, 51 studies on molecular entomology and 36 studies on molecular parasitology of malaria and three on human host were selected.

Results: Molecular methods are essential for improving the detection of malaria infection and monitoring antimalarial drugs and insecticide resistance in malaria elimination settings such as Iran.

Conclusion: The application of molecular methods may be of particular interest for malaria control/elimination programs, for monitoring progress towards malaria elimination, and for optimal orientation of program activities.

WHO. World malaria report 2017. World Health Organization. 2017

Gholizadeh S. Malaria zoonoses and its future challenges. J Bacteriol Parasitol. 2013;4:1000e1117.

White NJ. Plasmodium knowlesi: The fifth human malaria parasite. Clin Infect Dis. 2008; 46(2):172-3.

Brasil P, Zalis MG, de Pina-Costa A, Siqueira AM et al. Outbreak of human malaria caused by Plasmodium simium in the atlantic forest in rio de janeiro: A molecular epidemiological investigation. Lancet Glob Health. 2017; 5(10):e1038-e1046.

Latichev LN. Epidemic parasitology mission to Iran and parasitological surveys. Acad Sci USSR. 1948:235-238.

Jalaly Moslem Gh. History of malaria studies and malaria campaign in Iran. 1958; Resident thesis. Inst. Parasitol & Malariol. [In Persian]; 1958.

Raeisi A, Gouya MM, Nadim A et al. Determination of malaria epidemiological status in Iran's malarious areas as baseline information for implementation of malaria elimination program in Iran. Iran J Public Health. 2013; 42(3):326-33.

Nateghpour M, Jamshidi Makiani M, Mohseni G et al. First case of imported P. ovale from Iran. Iran J Med Sci. 2010;35:264-266.

Ghafari AN, Shahgudian ER. The Anopheles spp. of malaria vectors in Iran. In: Vector born diseases. 5th Medical Congress in Iran. . 1956

Gholizadeh S, Djadid ND, Nouroozi B, Bekmohammadi M. Molecular phylogenetic analysis of Anopheles and cellia subgenus anophelines (diptera: Culicidae) in temperate and tropical regions of Iran. Acta Trop. 2013; 126(1):63-74.

Edrissian GH. Malaria in Iran: Past and present situation. Iran J Parasitol. 2006;1:1-12.

Zaim M, Subbarao SK, Manouchehri AV, Cochrane AH. Role of Anopheles culicifacies s.L. and An. pulcherrimus in malaria transmission in Ghassreghand (Baluchistan), Iran. J Am Mosq Control Assoc. 1993; 9(1):23-6.

Azari-Hamidian S. Checklist of Iranian mosquitoes (diptera: Culicidae). J Vector Ecol. 2007; 32(2):235-42.

Zakeri S, Dinparast Djadid N, Zeinali S. Sequence heterogeneity of the merozoite surface protein-1 gene (msp-1) of Plasmodium vivax wild isolates in southeastern Iran. Acta Trop. 2003 ;88(1):91-7.

Ehtesham R, Fazaeli A, Raeisi A, Keshavarz H, Heidari A. Detection of mixed-species infections of Plasmodium falciparum and P. vivax by nested pcr and rapid diagnostic tests in southeastern Iran. Am J Trop Med Hyg. 2015; 93(1):181-5.

Khorashad SA, Salehi M, Roshanravan B. The comparison of microscopic method and rapid diagnostic test in detecting Plasmodium species. Int J Infect. 2014;1:e21441.

Li J, Wirtz RA, McCutchan TF. Analysis of malaria parasite rna from decade-old giemsa-stained blood smears and dried mosquitoes. Am J Trop Med Hyg. 1997; 57(6):727-31.

Okell LC, Ghani AC, Lyons E, Drakeley CJ. Submicroscopic infection in Plasmodium falciparum-endemic populations: A systematic review and meta-analysis. J Infect Dis. 2009; 200(10):1509-17.

Zakeri S, Najafabadi ST, Zare A, Djadid ND. Detection of malaria parasites by nested PCR in south-eastern, Iran: Evidence of highly mixed infections in chahbahar district. Malar J. 2002;1:2.

Zakeri S, Mamaghani S, Mehrizi AA, Shahsavari Z et al. Molecular evidence of mixed Plasmodium vivax and P. falciparum infections in northern islamic republic of Iran. East Mediterr Health J. 2004; 10(3):336-42.

Haghdoost AA, Mazhari S, Bahadini K. Comparing the results of light microscopy with the results of pcr method in the diagnosis of Plasmodium vivax. J Vector Borne Dis. 2006;43:53-57.

Ebrahimzadeh A, Fouladi B, Fazaeli A. High rate of detection of mixed infections of Plasmodium vivax and P. falciparum in south-east of Iran, using nested PCR. Parasitol Int. 2007; 56(1):61-4.

Nateghpour M, Khojasteh HA, Keshavarz H et al. Comparison of microscopical examination and semi-nested multiplex polymerase chain reaction in diagnosis of Plasmodium falciparum and P. vivax/comparai-son de l'examen microscopique et de l'amplification génique multiplex semi-nichée pour le diagnostic de P. falciparum et P. vivax. East Mediterr Health J. 2011;17:51.

Ghayour Najafabadi Z, Oormazdi H1, Akhlaghi L et al. Mitochondrial PCR-based malaria detection in saliva and urine of symptomatic patients. Trans R Soc Trop Med Hyg. 2014; 108(6):358-62.

Afsharpad M, Zakeri S, Pirahmadi S, Djadid ND. Molecular monitoring of Plasmodium falciparum resistance to antimalarial drugs after adoption of sulfadoxine-pyrimethamine plus artesunate as the first line treatment in Iran. Acta Trop. 2012; 121(1):13-8.

Ursing J, Zakeri S, Gil JP, Björkman A. Quinoline resistance associated polymorphisms in the pfcrt, pfmdr1 and pfmrp genes of Plasmodium falciparum in Iran. Acta Trop. 2006; 97(3):352-6.

Zakeri S, Afsharpad M, Kazemzadeh T et al. Association of pfcrt but not pfmdr1 alleles with chloroquine resistance in Iranian isolates of Plasmodium falciparum. Am J Trop Med Hyg. 2008; 78(4):633-40.

Zakeri S, Afsharpad M, Raeisi A, Djadid ND. Prevalence of mutations associated with antimalarial drugs in Plasmodium falciparum isolates prior to the introduction of sulphadoxine-pyrimethamine as first-line treatment in Iran. Malar J. 2007;6:148.

Eskandarian AA, Keshavarz H, Basco LK, Mahboudi F. Do mutations in Plasmodium falciparum dihydropteroate synthase and dihydrofolate reductase confer resistance to sulfadoxine-pyrimethamine in Iran? Trans R Soc Trop Med Hyg. 2002; 96(1):96-8.

Zakeri S, Farahani MS, Afsharpad M, Salehi M, Raeisi A, Djadid ND. High prevalence of the 437g mutation associated with sulfadoxine resistance among Plasmodium falciparum clinical isolates from Iran, three years after the introduction of sulfadoxine-pyrimethamine. Int J Infect Dis. 2010; 14 Suppl 3:e123-8.

Zakeri S, Gil JP, Bereckzy S, Djadid ND, Bjorkman A. High prevalence of double Plasmodium falciparum dhfr mutations at codons 108 and 59 in the sistan-baluchistan province, Iran. J Infect Dis. 2003; 187(11):1828-9.

Jafari S, Le Bras J, Asmar M, Durand R. Molecular survey of Plasmodium falciparum resistance in south-eastern Iran. Ann Trop Med Parasitol. 2003; 97(2):119-24.

Heidari A, Dittrich S, Jelinek T, Kheirandish A, Banihashemi K, Keshavarz H. Genotypes and in vivo resistance of Plasmodium falciparum isolates in an endemic region of Iran. Parasitol Res. 2007; 100(3):589-92.

Jalousian F, Dalimi A, Samiee SM, Ghaffarifar F, Soleymanloo F, Naghizadeh R. Mutation in pfmdr1 gene in chloroquine-resistant Plasmodium falciparum isolates, southeast Iran. Int J Infect Dis. 2008; 12(6):630-4.

Zakeri S1, Hemati S, Pirahmadi S, Afsharpad M, Raeisi A, Djadid ND. Molecular assessment of atpase6 mutations associated with artemisinin resistance among unexposed and exposed Plasmodium falciparum clinical isolates to artemisinin-based combination therapy. Malar J. 2012; 11:373.

Pirahmadi S, Zakeri S, Afsharpad M, Djadid ND. Mutation analysis in pfmdr1 and pfmrp1 as potential candidate genes for artemisinin resistance in Plasmodium falciparum clinical isolates 4 years after implementation of artemisinin combination therapy in Iran. Infect Genet Evol. 2013; 14:327-34.

Zakeri S, Motmaen SR, Afsharpad M, Djadid ND. Molecular characterization of antifolates resistance-associated genes, (dhfr and dhps) in Plasmodium vivax isolates from the middle east. Malar J. 2009;8:20.

Sharifi K, Haghighi A, Gachkar L et al. Molecular characterization of dihydrofolate reductase-thymidylatz synthase gene concerning antifulate resistance of P. vivax. Iran J Parasitol. 2009;4:10-18.

Afsharpad M, Zakeri S, Pirahmadi S, Djadid ND. Molecular assessment of dhfr/dhps mutations among P. vivax clinical isolates after introduction of sulfadoxine/pyrimethamine in combination with artesunate in Iran. Infect Genet Evol. 2012; 12(1):38-44.

Leroux-Roels G, Leroux-Roels I, Clement F et al. Evaluation of the immune response to rts,s/as01 and rts,s/as02 adjuvanted vaccines: Randomized, double-blind study in malaria-naive adults. Hum Vaccin Immunother. 2014; 10(8):2211-9.

Miura K, Jongert E, Deng B et al. Effect of ingested human antibodies induced by rts, s/as01 malaria vaccination in children on Plasmodium falciparum oocyst formation and sporogony in mosquitoes. Malar J. 2014;13:263.

Craig AA, Kain KC. Molecular analysis of strains of P. vivax from paired primary and relapse infections. J Infect Dis. 1996; 174(2):373-9.

Kolakovich KA, Ssengoba A, Wojcik K, Tsuboi T, al-Yaman F, Alpers M, Adams JH. P. vivax: Favored gene frequencies of the merozoite surface protein-1 and the multiplicity of infection in a malaria endemic region. Exp Parasitol. 1996; 83(1):11-9.

Miahipour A, Keshavarz H, Heidari A, Raeisi A, Rezaeian M, Rezaie S. Genetic variation of msp-1 gene in Plasmodium vivax isolated from patients in hormozgan province, Iran using sscp-PCR. Iran J Parasitol. 2012; 7(4):1-7.

Zakeri S, Barjesteh H, Djadid ND. Merozoite surface protein-3alpha is a reliable marker for population genetic analysis of Plasmodium vivax. Malar J. 2006 ;5:53.

Shahbazi A, Raeisi A, Nateghpour M et al. Polymorphism of merozoite surface protein-3α gene of P. vivax in isolates of Iran. Iran J Parasitol. 2008;3:15-20.

Shahbazi A, Mirhendi H, Raeisi A. P. vivax msp-3ß gene as a genetic marker for the parasite detection in comparison with ssrRNA gene. Iran J Publ Health. 2010;39:105-109.

Nateghpour M, Mavi SA, Keshavarz H, Rezaei S, Abedi F, Edrissian G, Raeisi A. Molecular monitoring of Plasmodium vivax infection after radical treatment in southeastern Iran. Iran J Arthropod Borne Dis. 2010; ;4(1):24-30.

Mehrizi AA, Zakeri S, Salmanian AH, Sanati MH, Djadid ND. Plasmodium falciparum: Sequence analysis of the gene encoding the c-terminus region of the merozoite surface protein-1, a potential malaria vaccine antigen, in Iranian clinical isolates. Exp Parasitol. 2008; 118(3):378-85.

Zamani Z, Razavi MR, Sadeghi S, Naddaf S et al. Sequence diversity of the c-terminal region of Plasmodium falciparum merozoite surface protein 1 in southern Iran. Southeast Asian J Trop Med Public Health. 2009; 40(1):1-9.

Heidari A, Keshavarz H, Rokni MB, Jelinek T. Genetic diversity in merozoite surface protein (msp)-1 and msp-2 genes of Plasmodium falciparum in a major endemic region of Iran. Korean J Parasitol. 2007; 45(1):59-63.

Heidari A, Keshavarz H, Dittrich S, Jelinek T. Allelic dimorphism of the Plasmodium falciparum erythrocyte binding antigen-175 (eba-175) gene in the south-east of Iran. Iran J Parasitol. 2009;4:17-22.

Zakeri S, Abouie Mehrizi A, Djadid ND, Snounou G. Circumsporozoite protein gene diversity among temperate and tropical Plasmodium vivax isolates from Iran. Trop Med Int Health. 2006; 11(5):729-37.

Cole-Tobian JL, Cortés A, Baisor M, Kastens W et al. Age-acquired immunity to a Plasmodium vivax invasion ligand, the duffy binding protein. J Infect Dis. 2002; 186(4):531-9.

Miller LH, Mason SJ, Clyde DF, McGinniss MH. The resistance factor to Plasmodium vivax in blacks. The duffy-bloodgroup genotype, fyfy. N Engl J Med. 1976; 295(6):302-4.

Babaeekho L, Zakeri S, Djadid ND. Genetic mapping of the duffy binding protein (dbp) ligand domain of Plasmodium vivax from unstable malaria region in the middle east. Am J Trop Med Hyg. 2009; 80(1):112-8.

Valizadeh V, Zakeri S, Mehrizi AA, Djadid ND. Population genetics and natural selection in the gene encoding the duffy binding protein II in Iranian Plasmodium vivax wild isolates. Infect Genet Evol. 2014; 21:424-35.

Mardani A, Keshavarz H, Heidari A, Hajjaran H, Raeisi A, Khorramizadeh MR. Genetic diversity and natural selection at the domain i of apical membrane antigen-1 (ama-1) of Plasmodium falciparum in isolates from Iran. Exp Parasitol. 2012 ;130(4):456-62.

Zakeri S, Sadeghi H, Mehrizi AA, Djadid ND. Population genetic structure and polymorphism analysis of gene encoding apical membrane antigen-1 (ama-1) of Iranian Plasmodium vivax wild isolates. Acta Trop. 2013; 126(3):269-79.

Motevalli Haghi A, Nateghpour M, Edrissian G et al. Sequence analysis of different domains of Plasmodium vivax apical membrane antigen (pvama-1 gene) locus in Iran. Iran J Parasitol. 2012; 7(1):26-31.

Djadid ND, Gholizadeh S, Aghajari M, Zehi AH, Raeisi A, Zakeri S. Genetic analysis of rDNA-ITS2 and rapd loci in field populations of the malaria vector, Anopheles stephensi (diptera: Culicidae): Implications for the control program in Iran. Acta Trop. 2006; 97(1):65-74.

Chavshin AR, Oshaghi MA, Vatandoost H et al. Molecular characterization, biological forms and sporozoite rate of Anopheles stephensi in southern Iran. Asian Pac J Trop Biomed. 2014;4:47-51.

Oshaghi MA, Yaaghoobi F, Abaie MR. Pattern of mitochondrial DNA variation between and within Anopheles stephensi (diptera: Culicidae) biological forms suggests extensive gene flow. Acta Trop. 2006; 99(2-3):226-33.

Gholizadeh S, Firooziyan S, Ladonni H et al. The Anopheles stephensi odorant binding protein 1 (asteobp1) gene, a new molecular marker for biological forms diagnosis. Acta Trop. 2015; 146:101-13.

Sedaghat MM, Linton YM, Oshaghi MA, Vatandoost H, Harbach RE. The Anopheles maculipennis complex (diptera: Culicidae) in Iran: Molecular characterization and recognition of a new species. Bull Entomol Res. 2003; 93(6):527-35.

Djadid ND, Gholizadeh S, Tafsiri E, Romi R, Gordeev M, Zakeri S. Molecular identification of palearctic members of Anopheles maculipennis in northern Iran. Malar J. 2007;6:6.

Djadid ND, Hassanzehi AG, Gholizadeh S. Studies on the role of a novel genetic variant of Anopheles culicifacies Giles, 1901 (Diptera: Culicidae) on malaria transmission in southeastern Iran. Ann Med Entomol. 2003;12:1-11.

Oshaghi MA, Taghilo B, Moradi MT, Vatandoost H. Detection of the Anopheles culicifacies complex species A and B in Baluchistan using mtDNA PCR-RFLP assay; the first report of species B from Iran. Hakim 2004;7:35-41.

Dezfouli SR, Oshaghi MA, Vatandoost H, Assmar M. Rdna-its2 based species-diagnostic polymerase chain reaction assay for identification of sibling species of Anopheles fluviatilis in Iran. Southeast Asian J Trop Med Public Health. 2003; 34 Suppl 2:56-60.

Chen B, Butlin RK, Pedro PM, Wang XZ, Harbach RE. Molecular variation, systematics and distribution of the Anopheles fluviatilis complex in southern Asia. Med Vet Entomol. 2006; 20(1):33-43.

Naddaf SR, Razavi MR, Bahramali G. Molecular variation and distribution of Anopheles fluviatilis (diptera: Culicidae) complex in Iran. Korean J Parasitol. 2010; 48(3):231-6.

Mehravaran A, Oshaghi MA, Vatandoost H, Abai MR, Ebrahimzadeh A, Roodi AM, Grouhi A. First report on Anopheles fluviatilis U in southeastern Iran. Acta Trop. 2011; 117(2):76-81.

Naddaf SR, Oshaghi MA, Vatandoost H. Confirmation of two sibling species among Anopheles fluviatilis mosquitoes in south and southeastern Iran by analysis of cytochrome oxidase I gene. J Arthropod Borne Dis. 2012; 6(2):144-50.

Oshaghi MA, Shemshad Kh, Yaghobi-Ershadi MR et al. Genetic structure of the malaria vector Anopheles superpictus in Iran using mitochondrial cytochrome oxidase (COI and COII) and morphologic markers: A new species complex? Acta Trop. 2007; 101(3):241-8.

Oshaghi MA, Yaghobi-Ershadi MR, Shemshad K et al. The Anopheles superpictus complex: Introduction of a new malaria vector complex in Iran. Bull Soc Pathol Exot. 2008; 101(5):429-34.

Oshaghi M, Moradi M, Taghilo B. Specific detection of malaria parasites using nested-PCR in individual mosquitoes and infected bloods in Chabahar and Iranshahr, Iran. Hakim. 2004;7:24-31.

Assmar M, Terhovanessian A, Naddaf SR et al. PCR detection of malaria parasites in Anopheles stephensi and An. culicifacies mosquitoes collected from southern endemic foci of Iran. J Sch Public Health Inst Public Health Res. 2005;3:19-26.

Djadid ND, Jazayeri H, Gholizadeh S, Rad ShP, Zakeri S. First record of a new member of Anopheles hyrcanus group from Iran: Molecular identification, diagnosis, phylogeny, status of kdr resistance and Plasmodium infection. J Med Entomol. 2009; 46(5):1084-93.

Gholizadeh S, Zakeri S, Djadid ND. Genotyping P. vivax isolates infecting Anopheles stephensi, an Asian main malaria vector. Exp Parasitol. 2013; 134(1):48-51.

Enayati AA, Vatandoost H, Ladonni H, Townson H, Hemingway J. Molecular evidence for a kdr-like pyrethroid resistance mechanism in the malaria vector mosquito An. stephensi. Med Vet Entomol. 2003; 17(2):138-44.

Djadid ND, Forouzesh F, Karimi M, Raeisi A et al. Monitoring pyrethroid insecticide resistance in major malaria vector Anopheles culicifacies: Comparison of molecular tools and conventional susceptibility test. Iran Biomed J. 2007; 11(3):169-176.

Djadid ND, Forouzesh F, Zakeri S. Identification of knock-down resistance (kdr) mutation in vgsc gene, related to pyrethroids resistance in Iranian Anopheles sacharovi and An. maculipennis. 4th Iranian Biotechnology Congress. 2005

Djadid ND, Barjesteh H, Raeisi A, Hassanzahi A, Zakeri S. Identification, sequence analysis, and comparative study on gste2 insecticide resistance gene in three main world malaria vectors: Anopheles stephensi, An. culicifacies, and An. fluviatilis. J Med Entomol. 2006; 43(6):1171-7.

Djadid ND, Barjesteh H, Forouzesh F, Zakeri S. Determination of glutathione s-transferase e2 region (gste2) in DDT susceptible and resistant Anopheles stephensi populations: Significance and application of nucleotide and amino acid comparison. Iran J Biotechnol. 2008;6:92-97.

Oshaghi MA, Chavshin AR, Vatandoost H, Yaaghoobi F, Mohtarami F, Noorjah N. Effects of post-ingestion and physical conditions on PCR amplification of host blood meal DNA in mosquitoes. Exp Parasitol. 2006; 112(4):232-6.

Oshaghi MA, Chavshin AR, Vatandoost H. Analysis of mosquito bloodmeals using rflp markers. Exp Parasitol. 2006; 114(4):259-64.

Anjomruz M, Oshaghi MA, Sedaghat MM et al. ABO blood groups of residents and the ABO host choice of malaria vectors in southern Iran. Exp Parasitol. 2014; 136:63-7.

IssueVol 13 No 2 (2018) QRcode
SectionReview Article(s)
Malaria Molecular methods Control Elimination Iran

Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
How to Cite
GHOLIZADEH S, NASERI KARIMI N, ZAKERI S, DINPARAST DJADID N. The Role of Molecular Techniques on Malaria Control and Elimination Programs in Iran: A Review Article. Iran J Parasitol. 2018;13(2):161-171.