Recombinant granule protein-1 for biomolecular diagnosis of toxoplasmosis in Indonesia

Wayan T. Artama, . and Sujono, . and Wiwien S. Utami, . (2009) Recombinant granule protein-1 for biomolecular diagnosis of toxoplasmosis in Indonesia. [Proceedings Paper]

Toxoplasmosis is a parasitic disease caused by intracellular parasite Toxoplasma gondii. Until now toxoplasmosis still is still a big problem in humans as well as animals in Indonesia. Human toxoplasmosis is dangerous if it inflicts immunodeficiency patients and pregnant women. Primary infection in pregnant women can cause abortion Intra Uterine Foetal Dead lUFD or born defect with hydrocephalus mental retardation or retinochorioditis. Toxoplsma gondii has a soluble protein or Excretory and Secretory Antigens ESAs that play an important role during active penetration of parasite into the cell target. Excretory and secretory antigens stimulates intracellular vacuole modification. Therefore after the parasite successfully enters the cell target the Granule GRA proteins are then responsible for the formation of parasitophorus vacuole which protect fusion with other intracellular compartments such as lysosomal vacuole. Consequently this parasite can survive and multiply at the cell target. The current study was aimed to subclone gra-1 gene in pET 32a and transformed those recombinant plasmid in E. coli in BL-21 in order to express GRA-1 recombinant protein. The recombinant protein is then purified by affinity chromatography on Nickel NTA and used for determining the prevalence of human toxoplasmosis in Yogyakarta using sandwich ELISA. The results of the study showed that the recombinant protein GRA-1 represented the ESAs protein for the determination of toxoplasmosis prevalence in animals and humans. As a preliminary results we found that the prevalence of toxoplasmosis in sheep was 49 goat 51 dairy cow 21 and Bali cattle 7 1 and in humans in five Districts of Yogyakarta as follows: Yogyakarta City 54.76; Sleman 60; Bantul 48.57; Gunung Kidul 20.48; and Kulon Progo 77.14.

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Abstract

Toxoplasmosis is a parasitic disease caused by intracellular parasite Toxoplasma gondii. Until now toxoplasmosis still is still a big problem in humans as well as animals in Indonesia. Human toxoplasmosis is dangerous if it inflicts immunodeficiency patients and pregnant women. Primary infection in pregnant women can cause abortion Intra Uterine Foetal Dead lUFD or born defect with hydrocephalus mental retardation or retinochorioditis. Toxoplsma gondii has a soluble protein or Excretory and Secretory Antigens ESAs that play an important role during active penetration of parasite into the cell target. Excretory and secretory antigens stimulates intracellular vacuole modification. Therefore after the parasite successfully enters the cell target the Granule GRA proteins are then responsible for the formation of parasitophorus vacuole which protect fusion with other intracellular compartments such as lysosomal vacuole. Consequently this parasite can survive and multiply at the cell target. The current study was aimed to subclone gra-1 gene in pET 32a and transformed those recombinant plasmid in E. coli in BL-21 in order to express GRA-1 recombinant protein. The recombinant protein is then purified by affinity chromatography on Nickel NTA and used for determining the prevalence of human toxoplasmosis in Yogyakarta using sandwich ELISA. The results of the study showed that the recombinant protein GRA-1 represented the ESAs protein for the determination of toxoplasmosis prevalence in animals and humans. As a preliminary results we found that the prevalence of toxoplasmosis in sheep was 49 goat 51 dairy cow 21 and Bali cattle 7 1 and in humans in five Districts of Yogyakarta as follows: Yogyakarta City 54.76; Sleman 60; Bantul 48.57; Gunung Kidul 20.48; and Kulon Progo 77.14.

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Item Type:	Proceedings Paper
Additional Information:	3ill. 19 ref.
AGROVOC Term:	Granules
AGROVOC Term:	Proteins
AGROVOC Term:	Biomolecular engineering
AGROVOC Term:	Toxoplasmosis
AGROVOC Term:	Indonesia
Geographical Term:	MALAYSIA
Depositing User:	Ms. Norfaezah Khomsan
Last Modified:	24 Apr 2025 05:27
URI:	http://webagris.upm.edu.my/id/eprint/15750

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