Evaluation of a Newly Designed Immunochromatographic Test using Gold Nanoparticles and Recombinant Antigen gra7 for Rapid Diagnosis of  Human Toxoplasmosis

Morovati, Hassan; Seyyedtabaei, Seyyed javad; Gholamzad, Mehrdad

doi:10.30699/ijmm.14.1.101

year 14, Issue 1 (January - February 2020) Iran J Med Microbiol 2020, 14(1): 101-115 | Back to browse issues page

‎ 10.30699/ijmm.14.1.101

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Morovati H, Seyyedtabaei S J, Gholamzad M. Evaluation of a Newly Designed Immunochromatographic Test using Gold Nanoparticles and Recombinant Antigen gra7 for Rapid Diagnosis of Human Toxoplasmosis. Iran J Med Microbiol 2020; 14 (1) :101-115
URL: http://ijmm.ir/article-1-1048-en.html

Evaluation of a Newly Designed Immunochromatographic Test using Gold Nanoparticles and Recombinant Antigen gra7 for Rapid Diagnosis of Human Toxoplasmosis

Hassan Morovati¹

, Seyyed javad Seyyedtabaei²

, Mehrdad Gholamzad

1- Department of Quality Control, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran
2- Department of Parasitology and Mycology, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
3- Department of Microbiology and Immunology, Faculty of Medicine, Tehran Medical Sciences, Islamic Azad University, Tehran, IRAN , mgholamzad@iautmu.ac.ir

Abstract: (5127 Views)

Background: One of the most important complications of toxoplasmosis is its early diagnosis. It seems that GRA7 protein can be a good candidate for detection of the acute phase in Toxoplasmosis. Accordingly, the present study aimed to diagnose toxoplasmosis via a newly immunochromatographic test using recombinant antigen gra7.
Methods: The parasite was cultured in mice and then were used for DNA extraction. The gra7 gene was amplified by PCR and cloned into the pET-32a (+) plasmid. Thereafter, the recombinant vector was transferred into the Escherichia coli Rosetta strain and gra7 was detected via SDS-PAGE and western blotting. The bacterial lysate was used to purify the protein by Ni-NTA affinity chromatography .Anti-human gold conjugated antibody, test line and control line were injected to conjugate pad and nitrocellulose membrane, respectively, and all the layer were assembled. By using serum of patients and healthy individuals, manufactured kits were evaluated.
Results: Our results indicated that the selected gene was correctly cloned and the protein of interest was produced and purified. The test revealed sensitivity and specificity of 100 and 96.7 percent, respectively. The kit was also shown to be stable over 16 weeks in 37°C.
Conclusion: The choice of antigen based on cellular and clinical features of the parasite, as well as the use of previous outcomes yielded to develop a rapid diagnostic test for toxoplasmosis.

Keywords: Toxoplasmosis, Immunochromatoga, gra7 antigen, RPD, Gold nanoparticles

Full-Text [PDF 1543 kb] (1763 Downloads) | | Full-Text (HTML) (1654 Views)

Type of Study: Original Research Article | Subject: Medical Parasitology
Received: 2019/11/27 | Accepted: 2019/12/29 | ePublished: 2020/01/1

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