Cloning and Sequence Analysis of Recombinant Plasmodium vivax Merozoite Surface Protein 1 (PvMSP-142 kDa) In pTZ57R/T Vector.

  • Hadi Mirahmadi Dept. of Parasitology and Mycology, Shahid Beheshti University of Medical Sciences, Tehran, Iran AND Infectious Diseases and Tropical Medicine Research Center, Zahedan University of Medical Sciences, Zahedan, Iran.
  • Adel Spotin Dept. of Parasitology and Mycology, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
  • Shirzad Fallahi Dept. of Parasitology and Mycology, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
  • Niloofar Taghipour Dept. of Parasitology and Mycology, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
  • Habibollah Turki Infectious and Tropical Diseases Research Center, Hormozgan University of Medical Sciences, Bandar Abbas, Iran.
  • Seyyed Javad Seyyed Tabaei Dept. of Parasitology and Mycology, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Keywords: Iran, Plasmodium vivax, Recombinant MSP-1 42 kDa, Sequencing

Abstract

Background:Haemonchosis has a negative effect on the farming industry throughout the world, especially in the tropic and sub-tropic countries. The present study was carried out to differentiate Haemonchus species from its main hosts in Iran, including sheep, goat and camel. Methods:The identification took place based on the morphometrics of the spic-ules and molecular characters. Two hundred seventy adult male nematodes were collected from the abomasums of different ruminants (90 samples from each ani-mal) at the slaughterhouses from different localities in Iran. Samples were morpho-logically identified according to the spicules’ morphometric measurements. In the section on molecular study, 10 samples of each Haemonchus isolates were genetically examined. A simple PCR-restriction fragment length polymorphism (PCR-RFLP) assay of the second internal transcribed spacer of ribosomal DNA (ITS2-rDNA) were described to confirm the PCR results.Results:PCR-RFLP profile obtained from the restriction enzyme HPa1 in H. con-tortus and H. longistipes indicated 1 (278 bp) and 2 (113 and 135 bp) different frag-ments, respectively. The morphological parameters clearly distinguish H. contortus from H. longistipes. Moreover, regarding the ITS2-rDNA, sequences of 295 bp and 314 bp were obtained from H. contortus and H. longistipes, respectively.Conclusion:High similarity of cloned PvMSP-142 kDa gene in comparison to reference sequence and other sequences could be beneficial as a remarkable mo-lecular marker for serological diagnostic.

References

Yeshiwondim AK, Tekle AH, Dengela DO, Yohannes AM, Teklehaimanot A. Therapeutic efficacy of chloroquine plus primaquine for threatment of Plasmodium vivax in Ethiopia. Ac-ta Trop. 2010; 113: 105-13.

Guerra CA, Howes RE, Patil AP, Gething PW, Van Moeckel TP, Temperly WH. The Interna-tional limits and population at risk of Plasmo-dium vivax transmission in 2009. PLoS Negl Trop Dis. 2010; 3: e744.

Arnott A, Barry AE, Reeder JC. Understanding the population genetics of Plasmodium vivax is essential for malaria control and elimination. Malar J. 2012; 11: 14.

Pasvol G. Management of severe malaria: inter-ventions and controversies. Infect Dis Clin North Am. 2005; 1: 211-40.

Figtree M, Passay C, Slade R, Cheng Q, Coolan N, Walker J, Saul A. Plasmodium vivax synony-mous frequencies, evolution and population structure deduced from diversity in AMA-1 and MSP-1 genes. Mol Biochem Parasitol. 2000; 108: 53-66.

Arevalo-Herrera M, Herrera S. Plasmodium vivax malaria vaccine development. Mol Immunol. 2001; 38: 443–55.

Blackman MJ, Scott-Finnigan TJ, Shai S, Hold-er AA. Antibodies inhibit the protease-medi-ated processing of a malaria merozoite surface protein. J Exp Med. 1994; 180: 389–93.

Uthaipibull C, Aufiero B, Syed SE et al. Inhibi-tory and blocking monoclonal antibody epitopes on merozoite surface protein 1 of the malaria parasite Plasmodium falciparum. J Mol Bi-ol. 2001; 13: 1381–94.

Mendis K, Sina BJ, Marchesini P, Carter R. The neglected burden of Plasmodium vivax ma-laria. Am J Trop Med Hyg. 2001; 64: 97–106.

Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S. MEGA5: Molecular Evolu-tionary Genetics Analysis using Maximum Likelihood, Evolutionary Distance, and Maxi-mum Parsimony Methods. Mol Biol Evol. 2011; 28: 2731-39.

Gerold P, Schofield L, Blackman MJ, Holder AA, Schwarz RT. Structural analysis of the gly-cosyl-phosphatidylinositol membrane anchor of the merozoite surface proteins-1 and -2 of Plasmodium falciparum. Mol Biochem Parasitol. 1996; 75: 131–43.

Holder AA, Guevara Patino JA, Uthaipibull C, Syed SE, Ling IT, Scott-Finnigan T, Blackman MJ. Merozoite surface protein 1, immune eva-sion, and vaccines against asexual blood stage malaria. Parasitologia. 1999; 41: 409–14.

Blackman MJ. Proteases involved in erythro-cyte invasion by the malaria parasite: function

and potential as chemotherapeutic targets. Curr Drug Targets. 2000; 1: 59 –83.

Blackman MJ, Whittle H, Holder AA. Pro-cessing of the Plasmodium falciparum major mer-ozoite surface protein-1: identification of a 33-kilodalton secondary processing product, which is shed prior to erythrocyte invasion. Mol Biochem Parasitol. 1991; 49: 35 – 44.

Odea KP, McKean PG, Harris A, Brown KN. Processing of the Plasmodium chabaudi AS mero-zoite surface protein 1 in vivo and in vitro. Mol Biochem Parasitol. 1995; 72: 111–9.

Tolle R, Fruh K, Doumbo O, Koita O, N’Diaye M, Fischer A, Dietz K, Bujard H. A prospective study of the association between the human humoral immune response to Plas-modium falciparum blood stage antigen gp190 and control of malarial infections. Infect Im-mun. 1993; 61: 40–7.

Pirson PJ, Perkins ME. Characterization with monoclonal antibodies of a surface antigen of Plasmodium falciparum merozoites. J Immunol. 1985; 134: 1946–51.

Chappel JA, Holder AA. Monoclonal antibod-ies that inhibit Plasmodium falciparum invasion in vitro recognize the first growth factor-like do-main of merozoite surface protein-1. Mol Bio-chem Parasitol. 1993; 60: 303–12.

Pacheco MA, Poe AC, Collins WE, Lal AA, Tanabe K, Kariuki SK, Udhayakumar V, Es-calante AA. A comparative study of the genetic diversity of the 42 kDa fragment of the mero-zoite surface protein 1 in P. falciparum and P. vi-vax. Infect Genet Evol. 2007; 7: 180–7.

Putaporntip C, Jongwutiwes S, Sakihama N, Ferreira MU, Kho WG, Kaneko A, et al., Mo-saic organization and heterogeneity in fre-quency of allelic recombination of the Plasmo-dium vivax merozoite surface protein-1 locus. Proc Natl Acad Sci USA. 2002; 99: 16348-53.

Walker-Abbey A, Djokam RR, Eno A, Leke RF, Titanji VP, Fogako J, Sama G, Thuita LH, Beardslee E, Snounou G, Zhou A, Taylor DW. Malaria in pregnant Cameroonian women: the effect of age and gravidity on submicroscopic and mixed species infections and multiple para-site genotypes. Am J Trop Med Hyg. 2005; 72: 229 –35.

Shahbazi A, Mirhendi H, Raeisi A. Plasmodium vivax MSP-3ß Gene as a Genetic Marker for the Parasite Detection in Comparison with Ssrrna Gene. Iran J Public Health. 2010; 39: 105-9.

Shahbazi A, Raeisi A, Nateghpour M, Mohe-bali M, Asmar M, Mirhendi H. Genetic struc-ture of Plasmodium vivax population assessed by sequence analysis of the merozoite surface pro-tein 3β gene. Iran J Clin Infect. 2010; 5: 126-32.

Zaman J, Shahbazi A, Asgharzadeh M. Plasmo-dium vivax dhfr Mutations among Isolates from

Malarious Areas of Iran. Korean J Parasitol. 2011; 49: 125-31.

Edrissian G. Malaria in Iran: past and Present Situation. Iran J Parasitol. 2006; 1: 1-14.

Shahbazi A, Farhadi P, Yerian M, Bazmani A, Khadem Nakhjiri S, Rasouli A, Raeisi A. De-tection of asymptomatic carriers of Plasmodium vivax among treated patients by Nested PCR method in Minab, Rudan and Bashagard, Iran. Iran J Parasitol. 2013; 8: 586-92.

How to Cite
1.
Mirahmadi H, Spotin A, Fallahi S, Taghipour N, Turki H, Seyyed Tabaei SJ. Cloning and Sequence Analysis of Recombinant Plasmodium vivax Merozoite Surface Protein 1 (PvMSP-142 kDa) In pTZ57R/T Vector. IJPA. 10(2):197-05.
Section
Original Article(s)