Abstract
Toxoplasma gondii infects all warm-blooded animals, including humans, causing serious public health problems and great economic loss for the food industry. Commonly used serological tests require costly and hazardous preparation of whole Toxoplasma lysate antigens from tachyzoites. Here, we have evaluated an alternative method for antigen production, which involved a prokaryotic expression system. Specifically, we expressed T. gondii dense granular protein-5 (GRA5) in Escherichia coli and isolated it by affinity purification. The serodiagnostic potential of the purified recombinant GRA5 (rGRA5) was tested through Western blot analysis against 212 human patient serum samples. We found that rGRA5 protein was 100% specific for analysis of toxoplasmosis-negative human sera. Also, rGRA5 was able to detect acute and chronic T. gondii infections (sensitivities of 46.8% and 61.2%, resp.).
Highlights
Toxoplasmosis is a parasitic disease caused by Toxoplasma gondii (T. gondii) which belongs to phylum Apicomplexa [1]
We polymerase chain reaction (PCR)-amplified a fragment of T. gondii granular protein-5 (GRA5) gene, which encoded amino acids 26–120 of the GRA5 protein
Upon induction of recombinant GRA5 (rGRA5) expression from GRA5-pRSET B-containing E. coli, we observed a 20 kDa band of increasing intensity, which was absent in the negative control
Summary
Toxoplasmosis is a parasitic disease caused by Toxoplasma gondii (T. gondii) which belongs to phylum Apicomplexa [1]. It is an obligate intracellular protozoan parasite capable of infecting all warm-blooded domestic animals as well as human beings [2]. Infection of T. gondii involves the transmission within and between hosts by zoites [4]. Three infectious stages of the parasite are tachyzoite, bradyzoite, and sporozoite [5]. Humans get infected with such disease through congenital transmission, consumption of raw or undercooked meat contaminated with T. gondii tissue cysts, or uptake of water contaminated with sporulated oocysts from the infected cat feces [6]
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