Designing Diagnostic Kit for Toxoplasma gondii Based on GRA7, SAG1, and ROP1 Antigens: An In Silico Strategy

et al. 2022

Sci Rep

Leishmaniasis is one of the main infectious diseases worldwide. In the midst of all the different forms of the disease, Cutaneous Leishmania (CL) has the highest incidence in the world. Many trial vaccines have been developed with the purpose of generating long-term cell-mediated immunity to Leishmania(L) major. As there is not any multi-epitope DNA vaccine with high efficacy against L.major, the aim of this study is to design a new multi-epitope DNA vaccine in order to have effective control upon this infectious disease through the immune bioinformatics. The L.major antigens: Gp63, LACK, TSA, LmSTI1and KMP11 were selected to design a multi-epitope DNA vaccine. The initial structure of the DNA vaccine was designed, benefiting from Gen Bank's website information. Epitopes of MHC-I antigens were predicted through the Immune Epitope Database (IEDB), and the selected epitopes were used to make vaccines construct along with linkers. New multi-epitope vaccine including 459 nucleic acids designed, and inserted between BamH1 and HindIII restriction sites of pCDNA3.1 mammalian expression vector. 12 epitopes among the chosen antigens were selected by two servers (IEDB and ANTIGEN). They had high stability and high antigenic power. Physicochemical features of vaccine measured by ProtParam server, and this structure was thermostable and hydrophilic. it’s a suitable model to study on the animal and human phases. The designed vaccine is expected to be an effective candidate through development of (CL) vaccines. However, the effectiveness of this vaccine should also evaluate in vivo model.

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Bioinformatics analysis for the purpose of designing a novel multi-epitope DNA vaccine against Leishmania major

Rashidi

Faraji

et al. 2022

Sci Rep

“…It seems that designing the most suitable immunogenic protein consisting of several immunogenic epitopes of two antigens S and N (proteins currently are being used in ELISA kits) is useful to increase the specificity and sensitivity of immunoassay-based tests. Having a multi-epitopic peptide that meets high specificity and sensitivity requirements that is a bottleneck in the immunoassay design processes facilitates development of rapid diagnostic tests with acceptable benefits (Aghamolaei et al 2020;Habibi et al 2020;Mamaghani et al 2020). Therefore, the aim of the present study is to predict and design a novel synthetic (fusion) protein consisting of multiple immune dominant B-cell epitopes from N and S proteins of SARS-CoV-2.…”

Section: Introductionmentioning

confidence: 99%

In-silico design of a multi-epitope for developing sero-diagnosis detection of SARS-CoV-2 using spike glycoprotein and nucleocapsid antigens

Netw Model Anal Health Inform Bioinforma

Arab-Mazar

Heidarzadeh

et al. 2021

Self Cite

COVID-19 is a pandemic disease caused by novel corona virus, SARS-CoV-2, initially originated from China. In response to this serious life-threatening disease, designing and developing more accurate and sensitive tests are crucial. The aim of this study is designing a multi-epitope of spike and nucleocapsid antigens of COVID-19 virus by bioinformatics methods. The sequences of nucleotides obtained from the NCBI Nucleotide Database. Transmembrane structures of proteins were predicted by TMHMM Server and the prediction of signal peptide of proteins was performed by Signal P Server. B-cell epitopes' prediction was performed by the online prediction server of IEDB server. Beta turn structure of linear epitopes was also performed using the IEDB server. Conformational epitope prediction was performed using the CBTOPE and eventually, eight antigenic epitopes with high physicochemical properties were selected, and then, all eight epitopes were blasted using the NCBI website. The analyses revealed that α-helices, extended strands, β-turns, and random coils were 28.59%, 23.25%, 3.38%, and 44.78% for S protein, 21.24%, 16.71%, 6.92%, and 55.13% for N Protein, respectively. The S and N protein three-dimensional structure was predicted using the prediction I-TASSER server. In the current study, bioinformatics tools were used to design a multi-epitope peptide based on the type of antigen and its physiochemical properties and SVM method (Machine Learning) to design multi-epitopes that have a high avidity against SARS-CoV-2 antibodies to detect infections by COVID-19.

“…Toxoplasma gondii is an obligate intracellular protozoan that causes infection in most warm‐blooded animals 1 . About one‐third of the world's population are infected with T .…”

Section: Introductionmentioning

confidence: 99%

“…Diagnostic approaches for detection of Toxoplasmosis include: serological assays, a variety of biological and molecular methods and histological and microscopic examinations but the specificity of diagnostic methods are not sufficient enough 1 . Also, microscopic and histological techniques have some technical errors in morphological detection like morphological similarity of diverse parasites in the same hosts, various microscopic forms of the same parasite in the different hosts, inappropriate staining, and incorrect recognition of tissue cyst from host 7–9 .…”

Section: Introductionmentioning

confidence: 99%

Immunodiagnosis and molecular validation of Toxoplasma gondii‐recombinant dense granular (GRA) 5 protein for the detection of toxoplasmosis in hemodialysis patients

Arab-Mazar

Fallahi

et al. 2021

Seminars in Dialysis

Self Cite

Toxoplasmosis causes serious complications in immunocompromised and pregnant women. Serological tests for the detection of toxoplasmosis are often designed from parasitic tachyzoites antigens. The process of producing these antigens is very difficult. The purpose of this study was evaluation of T. gondii‐rGRA5 for the immunodiagnosis and molecular detection of Toxoplasma infection using enzyme‐linked immunosorbent assay (ELISA) and LAMP methods in hemodialysis patients. The GRA5 gene was successfully expressed and purified by affinity chromatography assay and evaluated by western blot. Then it was used to design an ELISA assay. A total of 260 samples were tested for anti‐Toxoplasma IgG and IgM antibodies using a commercial ELISA kit and designed ELISA kit. Finally, the LAMP method was used to evaluate the precision and reliability of the results obtained by commercial and designed ELISA kits. The consistency of the results of two methods was analyzed using the Kappa coefficient of agreement. The rGRA5 revealed higher immunoreactivity with 1:100 dilution of sera from toxoplasmosis patients. The specificity and sensitivity of the assay were 93% and 96%, respectively. According to the Kappa coefficient, there was a substantial correlation between the results of ELISA and LAMP based on rGRA5 (≈98%, p < 0.001). Also it showed that rGRA5 protein can be used as an antigenic protein for designing sero‐diagnostic tests to identify Toxoplasma infection especially in hemodialysis patients.