Development of the multi-epitope chimeric antigen rqTSA-25 from Taenia saginata for serological diagnosis of bovine cysticercosis

Abstract

Bovine cysticercosis is a worldwide distributed zoonosis caused by the larval form of Taenia saginata present in bovine muscles. The diagnosis is based on the postmortem inspection at slaughterhouses and consists of the macroscopic visualization of lesions caused by cysticercosis in muscle sites. However, parasitized animals can pass unnoticed during sanitary inspection. Thus, the objective of this study was to characterize and evaluate the performance of different peptides from different regions of T. saginata for the cysticercosis diagnosis using enzyme-linked immunosorbent assay. We generated and evaluated a new recombinant protein chimera derived from the fusion of different peptides. We selected three distinct regions of T. saginata and predicted six peptides with antigenic potential (EP2–EP7). These peptides were analyzed individually and selected for generating a new chimeric recombinant protein. The new protein was termed rqTSA-25, and its performance rates were: 93.3% sensitivity (confidence interval (CI) = 76–98%), 95.3% specificity (CI = 82–99%), 93% positive predictive value (CI = 76–98%), 95% negative predictive value (CI = 82–99%), and 95% accuracy. In the immunoblot, this protein showed no false positive or false negative reaction. Thus, the use of rqTSA-25 is recommended for the diagnosis of bovine cysticercosis.

Systematic review and meta-analysis of bovine cysticercosis in Brazil: current knowledge and way forward

“…In addition, we found 12 articles containing information about the spatial distribution of BCC inside/within the states (Table 1), 13 articles describing risk factors or variables associated with BCC occurrence ( Table 2) and 4 describing the economic burden of BCC [7,22,28,30]. Furthermore, a set of 10 studies performed in Brazil were also included, regarding at least one aspect presented in discussion section, such as efficacy of chemical treatment (n = 1) [62], preferential infection sites (n = 2) [63,64], development of serological tests (n = 4) [65][66][67][68] and measures for BCC control (n = 3) [69][70][71].…”

“…The use of serological analyses, such as Ag-ELISA, to detect infected animals also has been suggested but might not be feasible during slaughter [12]. However, serological analyses have been largely carried out, including in Brazil [65][66][67][68] and its use to detect infected animals has improved worldwide [90,95,96], allowing to perform well-designed epidemiological studies.…”

Recombinant multiepitope proteins expressed in Escherichia coli cells and their potential for immunodiagnosis

“…coli cells ELISA 114 schistosomiasis-positive serum samples/92 healthy buffalo serum samples/14 serum samples from non-schistosomiasis diseases rBSjPGM-BSjRAD23-1-BSj23 Sensitivity: 95.61% Specificity: 97.83% rBSjRAD23-2-BSjPGM-BSj23 Sensitivity: 67.54% Specificity: 100% [ 121 ]/China Bovine cysticercosis/ Taenia saginata rqTSA-25 E . coli BL21-Codon-Plus(DE3)-RIL cells ELISA/immunoblot assay 60 cysticercosis-positive serum samples/30 healthy bovine serum samples/15 serum samples from non-cysticercosis diseases ELISA Sensitivity: 93.3% Specificity: 95.3% Immunoblot no false positive or false negative was observed [ 122 ]/Brazil Sheep cystic echinococcosis/ Echinococcus granulosus reEg mefAg-1 E. coli BL21(DE3) cells Indirect ELISA 86 echinococcosis-positive serum samples/30 echinococcosis-negative serum samples/21 serum samples from non- Echinococcus granulosus diseases Sensitivity: 93.41% Specificity: 99.31% [ 123 ]/China Human onchocerciasis/ Onchocerca volvulus OvNMP-48 – ELISA 101 onchocerciasis-positive serum samples/58 serum samples from healthy individuals/54 serum samples from non-onchocerciasis diseases Sensitivity: 76.0% Specificity: 97.4% [ 124 ]/Belgium Human fascioliasis/ Fasciola hepatica rMEP E . coli BL21 cells Western blot – rMEP was recognized by human positive samples [ 125 ]/Iran Human lymphatic filariasis/ Wuchereria bancrofti Multiepitope antigen E .…”

“…This lineage harbors extra copies of genes encoding rare tRNAs, including for AUA, AGG, AGA, CUA, CCC, and GGA codons [ 142 ]. The strain BL21-CodonPlus(DE3)-RIL, which contains similar modifications, was also used to express the RMP rqTSA-25 for the diagnosis of bovine tapeworm [ 122 ]. These examples demonstrate the ability to address codon usage issues by selecting suitable lineages without entirely replacing the gene codons planning.…”

Chimeric Protein Designed by Genome-Scale Immunoinformatics Enhances Serodiagnosis of Bovine Neosporosis

“…Recombinant antigens are produced on a large scale and are standardized for diagnostic tests to decrease the risks of cross-reaction (16). The use of chimeric proteins increases in an attempt to enhance diagnostic power in a combination of multiple antigens (20). In addition, synthetic antigens are easily manufactured, reaching high levels of purity in the absence of any living organism (21).…”