Production of refolded Toxoplasma gondii recombinant SAG1-related sequence 3 (SRS3) and its use for serodiagnosis of human toxoplasmosis

Mirzadeh, Abolfazl; Saadatnia, Geita; Golkar, Majid; Babaie, Jalal; Noordin, Rahmah

doi:10.1016/j.pep.2017.03.001

Cited by 12 publications

(5 citation statements)

References 38 publications

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“…In the past two decades, several Toxoplasma recombinant antigens have been cloned and expressed in E. coli or yeast for use as diagnostic reagents for human and animal infections. They include antigens from the parasite surface (SAGs), matrix (MAGs), dense granule (GRAs), rhoptry (ROPs), and microneme (MICs) [43][44][45][46][47][48][49][50]. Four of the recombinant antigens (SAG1, SAG2, GRA3, and GRA6) expressed in E. coli have been used in an RDT to detect infection in cats.…”

Section: Lateral Flow Assaymentioning

confidence: 99%

Serological and molecular rapid diagnostic tests for Toxoplasma infection in humans and animals

Khan

Noordin

2019

Eur J Clin Microbiol Infect Dis

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Infection by Toxoplasma gondii is prevalent worldwide. The parasite can infect a broad spectrum of vertebrate hosts, but infection of fetuses and immunocompromised patients is of particular concern. Easy-to-perform, robust, and highly sensitive and specific methods to detect Toxoplasma infection are important for the treatment and management of patients. Rapid diagnostic methods that do not sacrifice the accuracy of the assay and give reproducible results in a short time are highly desirable. In this context, rapid diagnostic tests (RDTs), especially with point-of-care (POC) features, are promising diagnostic methods in clinical microbiology laboratories, especially in areas with minimal laboratory facilities. More advanced methods using microfluidics and sensor technology will be the future trend. In this review, we discuss serological and molecular-based rapid diagnostic tests for detecting Toxoplasma infection in humans as well as animals.

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Section: Lateral Flow Assaymentioning

confidence: 99%

Serological and molecular rapid diagnostic tests for Toxoplasma infection in humans and animals

Khan

Noordin

2019

Eur J Clin Microbiol Infect Dis

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“…In addition, mature SRS3 and SAG1 contain 18). These findings suggest that they have similar folding, common overall topology, and the same function in stimulating the host system (16,18,19).…”

Section: Introductionmentioning

confidence: 71%

In Silico Prediction of T and B Cell Epitopes of SAG1-Related Sequence 3 (SRS3) Gene for Developing Toxoplasma gondii Vaccine

Mirzadeh

Saadatnia

Golkar

et al. 2020

Arch Clin Infect Dis

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: Toxoplasmosis is a worldwide infection that can lead to serious problems in immune-compromised individuals and fetuses. A DNA vaccine strategy would be an ideal tool against Toxoplasma gondii. One of the necessary measures to provide an effective vaccine is the selection of proteins with high antigenicity. The SAG1-related sequence 3 (SRS3) protein is a major surface antigen in T. gondii that can be used as a vaccine candidate. In the present study, bioinformatics and computational methods were utilized to predict protein characteristics, as well as secondary and tertiary structures. The in silico approach is highly suited to analyze, design, and evaluate DNA vaccine strategies. Hence, in silico prediction was used to identify B and T cell epitopes and compare the antigenicity of SRS3 and other candidate genes of Toxoplasma previously applied in the production of vaccines. The results of the analysis theoretically showed that SRS3 has multiple epitopes with high antigenicity, proposing that SRS3 is a promising immunogenic candidate for the development of DNA vaccines against toxoplasmosis.

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“…Furthermore, it has been shown that obtaining large quantities of highly pure recombinant F . hepatica antigens with correct folding play a vital role in improvement of antigenicity and accuracy in serodiagnosis methods [ 14 , 27 ]. In this line, we evaluated the immunodiagnostic potential of tFhSAP-2, tFh-Ftn-1 and FhLAP recombinant antigens, alone or in combination, for the detection of IgG antibodies in fascioliasis by ELISA.…”

Section: Discussionmentioning

confidence: 99%

“…coli antigen as a control, expressed and purified in the same way as the F . hepatica recombinant antigens [ 27 ]. The folded recombinant proteins alone and in combination was compared to E/S antigen in ELISA test, it was illustrated that the sensitivity of antibody diagnosis obtained from the combined antigens was the same as E/S antigen at 96% and slightly more than rtFhSAP-2 (95%).…”

Section: Discussionmentioning

confidence: 99%

“…Analysis of recombinant proteins solubility was carried out in E . coli strain as described previously [ 27 ]. One ml of each induced culture was pelleted and the crude cell extract was obtained in 10 ml of extraction buffer (0.1 mM EDTA, 1mM dithiothreitol (DTT), 30 mM Tris–HCl, pH 8 and then the mix was shaken in the cold room for 30 min.…”

Section: Bacterial Strains Plasmidsmentioning

confidence: 99%

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Evaluation of a set of refolded recombinant antigens for serodiagnosis of human fascioliasis

et al. 2018

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Diagnosis of fascioliasis with high sensitivity and specificity antigens play a vital role in the management of the disease. Majority of commercial serological tests use F. hepatica native antigens and indicate wide diversities in test accuracy. Nowadays, recombinant antigens have been introduced as diagnostic reagents offer better test standardization. A combination of highly pure recombinant antigens associated with correct folding will leads to improve specificity and sensitivity of ELISA for diagnosis of Fascioliasis. In this article, Fasciola hepatica saposin-like protein 2 (SAP-2), ferritin protein (Ftn-1) and leucine aminopeptidase (LAP) recombinant antigens were considered as tools for the detection of F. hepatica immunoglobulin G antibodies in persons with chronic human fasciolasis. The recombinant antigens were obtained as fusion proteins, expressed in Escherichia coli and purified by immobilized metal affinity chromatography (IMAC). The refolding processes of denatured recombinant proteins were performed using dialysis method in the presence of chemical additives, and reduced/oxidized glutathione (in vitro). The immunoreactivity of the recombinant antigens was assessed individually and in a combination compared with excretory/secretory antigen (E/S) in an enzyme-linked immunosorbent assay (ELISA) test. The experiments were optimized using 213 serum samples from humans, including patients with chronic fascioliasis, patients with other parasitic diseases, and healthy subjects. The results indicated 95% sensitivity and 98% specificity for rtFhSAP-2, 96% sensitivity and 91% specificity for E/S, 80% and 83.3% for rtFhFtn-1, 84% and 88% for FhLAP, and also, 96% and 95% for combination of recombinant antigens, respectively. In conclusion, the results of this investigation showed that rtFhSAP-2 with the highest specificity and acceptable sensitivity has a considerable superiority compared to mentioned antigens and even in combination with these antigens in serodiagnosis of human fascioliasis.

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Production of refolded Toxoplasma gondii recombinant SAG1-related sequence 3 (SRS3) and its use for serodiagnosis of human toxoplasmosis

Cited by 12 publications

References 38 publications

Serological and molecular rapid diagnostic tests for Toxoplasma infection in humans and animals

Serological and molecular rapid diagnostic tests for Toxoplasma infection in humans and animals

In Silico Prediction of T and B Cell Epitopes of SAG1-Related Sequence 3 (SRS3) Gene for Developing Toxoplasma gondii Vaccine

Evaluation of a set of refolded recombinant antigens for serodiagnosis of human fascioliasis

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