Detection of Asymptomatic Carriers of Plasmodium vivax among Treated Patients by Nested PCR Method in Minab, Rudan and Bashagard, Iran.

  • Abbas Shahbazi Tabriz Research Center of Infectious and Tropical Diseases, Tabriz University of Medical Sciences, Tabriz, Iran.
  • Pegah Farhadi Dept. of Medical Parasitology and Mycology, Tabriz University of Medical Sciences, Tabriz, Iran.
  • Masoud Yerian Institute of Public Health Researches, Tehran University of Medical Sciences, Tehran, Iran.
  • Ahad Bazmani Tabriz Research Center of Infectious and Tropical Diseases, Tabriz University of Medical Sciences, Tabriz, Iran.
  • Sara Khadem Nakhjiri School of Medicine, Gorgan University of Medical Sciences, Gorgan, Iran.
  • Arash Rasouli Dept. of Medical Parasitology and Mycology, Tabriz University of Medical Sciences, Tabriz, Iran.
  • Ahmad Raeisi Institute of Public Health Researches, Tehran University of Medical Sciences, Disease Management Center, Ministry of Health, Iran.
Keywords: Asymptomatic, Iran, PCR, Plasmodium vivax


 Background: Plasmodium vivax is the most widespread species of Plasmodium in humans and causing about 80 million clinical cases annually. This study was under-taken to detect P. vivax in asymptomatic treated vivax malaria patients to trace la-tent/sub-patent malaria infection. Method: The venous blood of all detected cases with P. vivax in Bashagard, Minab and Roodan Districts in Hormozgan Province from 2009 to 2010 was examined by microscopic and nested PCR methods for presence of the parasite. Results: In microscopic examination of peripheral blood smears, all samples were negative for the presence of the parasites. But, we detected two P. vivax related bands in the electrophoresis of the nested PCR products (120 bp). Conclusion: Following up the malaria cases after treatment by a combination of methods, or new diagnostics such as RDTs can be included in the priorities of ma-laria elimination program in Iran.


Suárez-Mutis MC, Cuervo P, Leoratti FM, Moraes-Avila SL, Ferreira AW, Fernandes O, Coura JR. Cross sectional study reveals a high percentage of asymptomatic Plasmodium vivax infection in the Amazon Rio Negro area, Brazil. Rev Inst Med Trop Sao Paulo. 2007; 49(3):159-64.

Harris I, Sharrock WW, Bain LM, Gray KA, Bobogare A, Boaz L, Lilley K, Krause D, Vallely A, Johnson ML, Gatton ML,Shanks GD, Cheng Q. A large proportion of asymp-tomatic Plasmodium infections with low and sub microscopic parasite densities in the low transmission setting of Temotu Province, Solomon Islands: challenges for malaria di-agnostics in anelimination setting. Malar J. 2010; 9:254.

Morgan, U. M. & Thompson, R. C. A. Mo-lecular detection of parasitic protozoa. Para-sitology. 1998; 117: S73-S85.

Zaman J, Shahbazi A, Asgharzadeh M. Plass-modium vivax dhfr mutations in isolates from malarious areas of Iran. Korean J Parasitol. 2011; 49( 2):125-131.

Zeyrek YF, Kurcer MA, Zeyrek D, Simsek Z. Parasite density and serum cytokine levels in Plasmodium vivax malaria in Turkey. Parasite Immunol. 2006; 28:201–207.

Sundar S, Rai M. Laboratory diagnosis of vis-ceral leishmaniasis. Clin Diagn Lab Immunol. 2002; 9:951–958.

Boonma P, Christensen PR, Suwanarusk R, Price RN, Russell B, Lek-Uthai U. Compari-son of three molecular methods for the de-tection and speciation of Plasmodium vivax and Plasmodium falciparum. Malar J. 2007; 6:124.

Schneider P, Wolters L, Schoone G, Schallig H, Sillekens P, Hermsen R, Sauerwein R. Re-al-time nucleic acid sequence-based ampli-fication is more convenient than real-time PCR for quantification of Plasmodium falcipa-rum. J Clin Microbiol. 2005; 43:402–405.

veron V, Simon S, Carme B. Multiplex real-time PCR detection of Plasmodium falciparum, P. vivax and P. malariae in human blood sam-ples. Exp Parasitol. 2009;121:346–351.

Snounou G, Viriyakosol S, Zhu XP, Jarra W, Pinheiro L, do Rosario VE, Thaithong S, Brown KN. High sensitivity of detection of human malaria parasites by the use of nested polymerase chain reaction. Mol Biochem Parasitol. 1993;61:315–320.

Gama BE, Silva-PiresFdo E, Lopes MN, Cardoso MA, Britto C, Torres KL, de Men-donça Lima L, de Souza JM, Daniel-Ribeiro CT, Ferreira-da-Cruz Mde F. Real-time PCR versus conventional PCR for malaria parasite detection in low-grade parasitemia. Exp Para-sitol. 2007;116:427–432.

McKenzie FE, Sirichaisinthop J, Miller RS, Gasser RA, Jr, Wongsrichanalai C. Depend-ence of malaria detection and species diagno-sis by microscopy on parasite density. Am J Trop Med Hyg. 2003;69:372–376.

Safeukui I, Millet P, Boucher S, Melinard L, Fregeville F, Receveur MC, Pistone T, Fialon P, Vincendeau P, Fleury H, Malvy D. Evalua-tion of FRET real-time PCR assay for rapid detection and differentiation of Plasmodium species in returning travelers and migrants. Malar J. 2008;7:70.

Babiker HA, Schneider P, Reece SE: Game-tocytes: insights gained during a decade of molecular monitoring. Trends Parasitol 2008, 24:525-530.

Farcas GA, Zhong KJ, Mazzulli T, Kain KC. Evaluation of the Real-Art Malaria LC real-time PCR assay for malaria diagnosis. J Clin Microbiol. 2004;42:636–638.

Andrews L, Andersen RF, Webster D, Du-nachie S, Walther RM, Bejon P, Hunt-Cooke A, Bergson G, Sanderson F, Hill AV, Gilbert SC. Quantitative real-time polymerase chain reaction for malaria diagnosis and its use in malaria vaccine clinical trials. Am J Trop Med Hyg. 2005;73:191–198.

Elsayed S, Plewes K, Church D, Chow B, Zhang K. Use of molecular beacon probes for real-time PCR detection of Plasmodium fal-ciparum and other Plasmodium species in pe-ripheral blood specimens. J Clin Microbiol. 2006;44:622–624.

Vo TK, Bigot P, Gazin P, Sinou V, De Pina JJ, Huynh DC, Fumoux F, Parzy D. Evalua-tion of a real-time PCR assay for malaria di-agnosis in patients from Vietnam and in re-turned travelers. Trans R Soc Trop Med Hyg. 2007;101:422–428

Tada MS, Marques RP, Mesquita E, Dalla Martha RC, Rodrigues JA, Costa JD, et al. Urban malaria in the Brazilian Western Ama-zon Region I: high prevalence of asympto-matic carriers in an urban riverside district is associated with a high level of clinical malaria. Mem Inst Oswaldo Cruz. 2007;102(3):263-269.

Alves FP, Gil lh, Marrelli MT, Ribolla PE, Camargo EP, Da Silva LH. Asymptomatic carriers of Plasmodium spp. As infection source for malaria vector mosquitoes in the Brazilian Amazon. J Med Entomol. 2005;42(5):777-779.

Haghdoost AA, Mazhari S, Bahaadini K. Es-timating the relapse risk of Plasmodium vivax in Iran under national chemotherapy scheme using a novel method. J Vector Borne Dis. 2006;43(4):168-172.

Diseases Management Center of MOH, I.R. Iran.Annual Reports of Malaria. 2006.

Roper C, Elhassan IM, Hviid L, Giha H, Richardson W, Babiker H, Satti GM, Thean-der TG, Arnot DE. Detection of very low level Plasmodium falciparum infections using the nested polymerase chain reaction and a reas-sessment of the epidemiology of unstable malaria in Sudan. Am J Trop Med Hyg. 1996;54:325-331.

Alves FP, Durlacher RR, MenezesMJ,Krieger H, Silva LH, Camargo EP. High prevalence of asymptomatic Plasmodium vivax and Plasmo-dium falciparum infections in native Amazoni-an populations. Am J Trop Med Hyg. 2002; 66(6):641-8.

Turki H, Zoghi S, Mehrizi AA, Zakeri S, Raeisi A, Khazan H, Haghdoost AA. Ab-sence of asymptomatic malaria infection in endemic area of Bashagard District, Hor-mozgan Province, Iran Iranian J Parasitol: Vol. 7, No.1, 2012; pp.36-44.

Shahbazi A, Raeisi A, Asmar M, Naddaf S, Nateghpour M, Anaraki G. Detection of Plasmodium vivax by Nested PCR in malari-ous areas of western north of Iran. Bi-monthly Journal of Hormozgan University of Medical Sciences .3. 2009; 13 (1) :7-12 (In Persian).

Zakeri S, Najafabadi TS, Zare A and Dinpar-ast N D. Detection of malaria parasites by nested PCR in south-eastern, Iran: Evidence of highly mixed infections in Chahbahar dis-trict. Malaria J. 2002;1:2.

Coleman RE, Sattabongkot J, PromstapormS, Maneechai N, Tippayachai B, KengluechaA, Rachapaew N, Zollner G, Miller RS, Vaughan JA, Thimasarn K, Khuntirat B. Comparison of PCR and microscopy for the detection of asymptomatic malaria in a Plas-modium falciparum/vivax endemic area in Thai-land. Malar J. 2006; 5: 121.

Zulma Milena Cucunubá, Ángela Patricia Guerra, Sonia Judith Rahirant, Jorge Alonso Rivera, Liliana Jazmín Cortés, Rubén Santia-go Nicholls. Asymptomatic Plasmodium spp. infection in Tierralta, Colombia. Mem Inst Oswaldo Cruz, Rio de Janeiro. November 2008; 103(7): 668-673.

Zakeri S, Kakar Q, Ghasemi F, Raeisi A, Butt W, Safi N, Afsharpad M, Memon S M, Gholizadeh S, Salehi M, Atta H, Zamanin Gh. Dinparast DN. Detection of mixed Plasmodium falciparum & P. vivax infections by nested-PCR in Pakistan, Iran & Afghanistan. Indian J Med Res. 2010;132: 31-35.

Shahbazi A, Raeisi A, Nateghpour M, Mir-hendi H, Mohebali M, Asmar M. Polymorphism of merozoite surface protein-3α gene of Plasmodium vivax in isolates of Iran. Iranian J Parasitol. 2008;3(2):15-20.

Shahbazi A, Raeisi A, Nateghpour M, Mohe-bali M, Asmar M, Mirhendi H. Partial Se-quence Analysis of Merozoite Surface Pro-teine -3α Gene in Plasmodium vivax isolates from malarious areas of Iran. Iranian J Parasi-tol. 2008;3(4):1-8.

Shahbazi A, Mirhendi H, Raeisi A. Plasmodium vivax MSP-3ß gene as a genetic marker for the parasite detection in comparison with ssrrna gene. Iranian J Publ Health. 2010; 39(2):105-109.

Nateghpour M, Akbarzadeh K, Farivar L, Amiri A. Detection of asymptomatic malaria infection among the Afghani immigrant pop-ulation in Iranshahr district of southeastern Iran. Bull Soc Pathol Exot. 2011 Oct;104(4):321-3.

How to Cite
Shahbazi A, Farhadi P, Yerian M, Bazmani A, Nakhjiri SK, Rasouli A, Raeisi A. Detection of Asymptomatic Carriers of Plasmodium vivax among Treated Patients by Nested PCR Method in Minab, Rudan and Bashagard, Iran. IJPA. 8(4):586-92.