Preliminary Evaluation of Recombinant EPC1 and TPx for Serological Diagnosis of Animal Cystic Echinococcosis

Liu, Yuqing; Song, Hai-Liang; Wu, Maodi; Xie, Yue; Gu, Xiaobin; He, Ran; Lai, Weiming; Jing, Bo; Peng, Xuerong; Yang, Guangyou

doi:10.3389/fcimb.2020.00177

Cited by 7 publications

(10 citation statements)

References 42 publications

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“…At the individual level, sensitivity and specificity were estimated at 55% and 68% respectively (for an OD cut-off value of 0.378). It is well known that the diagnostic performance of E. granulosus ELISA assays is highly variable, with previously published sensitivity and specificity values ranging from 41% to 100% [ 25 , 26 , 28 ]. Variation in individual immune responses, cyst location, and antigen characteristics have been linked to variable ELISA sensitivity and specificity.…”

Section: Discussionmentioning

confidence: 99%

“…These techniques can be used to detect host antibodies specific to E. granulosus in serum samples, allowing for the detection of exposed/infected flocks without the need for necropsy. However, despite improvements to these approaches through the use of recombinant antigens, their diagnostic performance can be highly variable in the field, limiting their effectiveness as surveillance tools [ 23 , [25] , [26] , [27] , [28] ].…”

Section: Introductionmentioning

confidence: 99%

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Modelling diagnostics for Echinococcus granulosus surveillance in sheep using Latent Class Analysis: Argentina as a case study

et al. 2022

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Echinococcus granulosus sensu lato is a globally prevalent zoonotic parasitic cestode leading to cystic echinococcosis (CE) in both humans and sheep with both medical and financial impacts, whose reduction requires the application of a One Health approach to its control. Regarding the animal health component of this approach, lack of accurate and practical diagnostics in livestock impedes the assessment of disease burden and the implementation and evaluation of control strategies. We use of a Bayesian Latent Class Analysis (LCA) model to estimate ovine CE prevalence in sheep samples from the Río Negro province of Argentina accounting for uncertainty in the diagnostics. We use model outputs to evaluate the performance of a novel recombinant B8/2 antigen B subunit (rEgAgB8/2) indirect enzyme-linked immunosorbent assay (ELISA) for detecting E. granulosus in sheep. Necropsy (as a partial gold standard), western blot (WB) and ELISA diagnostic data were collected from 79 sheep within two Río Negro slaughterhouses, and used to estimate individual infection status (assigned as a latent variable within the model). Using the model outputs, the performance of the novel ELISA at both individual and flock levels was evaluated, respectively, using a receiver operating characteristic (ROC) curve, and simulating a range of sample sizes and prevalence levels within hypothetical flocks. The estimated (mean) prevalence of ovine CE was 27.5% (95%Bayesian credible interval (95%BCI): 13.8%–58.9%) within the sample population. At the individual level, the ELISA had a mean sensitivity and specificity of 55% (95%BCI: 46%–68%) and 68% (95%BCI: 63%–92%), respectively, at an optimal optical density (OD) threshold of 0.378. At the flock level, the ELISA had an 80% probability of correctly classifying infection at an optimal cut-off threshold of 0.496. These results suggest that the novel ELISA could play a useful role as a flock-level diagnostic for CE surveillance in the region, supplementing surveillance activities in the human population and thus strengthening a One Health approach. Importantly, selection of ELISA cut-off threshold values must be tailored according to the epidemiological situation.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Modelling diagnostics for Echinococcus granulosus surveillance in sheep using Latent Class Analysis: Argentina as a case study

et al. 2022

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“…12 As a diagnostic antigen, it has high sensitivity and specificity. 13 The pET28a-P29/BL21 strain, which was constructed and preserved in our laboratory, was sequenced, confirmed to be free of mutation, and purified by nickel column affinity chromatography using isopropyl-β-Dthiogalactoside (IPTG)-induced expression to obtain the recombinant protein of E. granulosus P29 (rEg. P29).…”

Section: Introductionmentioning

confidence: 99%

“…In addition, other novel vaccines are being developed, including those based on the P29 protein, a new 29 kDa antigen first identified by Gonzalez et al from the protocephalic nodes of E. granulosus 12 . As a diagnostic antigen, it has high sensitivity and specificity 13 . The pET28a‐P29/BL21 strain, which was constructed and preserved in our laboratory, was sequenced, confirmed to be free of mutation, and purified by nickel column affinity chromatography using isopropyl‐β‐ d ‐thiogalactoside (IPTG)‐induced expression to obtain the recombinant protein of E. granulosus P29 (rEg.…”

Section: Introductionmentioning

confidence: 99%

Changes in intestinal flora of mice induced by rEg.P29 epitope peptide vaccines

Zhang,

Lv,

Zhao

et al. 2023

Immunity Inflam & Disease

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ObjectiveCystic echinococcosis (CE), a zoonotic parasitic disease caused by Echinococcus granulosus, remains a public health and socioeconomic issue worldwide, making its prevention and treatment of vital importance. The aim of this study was to investigate changes in the intestinal microbiota of mice immunized with three peptide vaccines based on the recombinant antigen of E. granulosus, P29 (rEg.P29), with the hope of providing more valuable information for the development of vaccines against CE.MethodsThree peptide vaccines, rEg.P29T, rEg.P29B, and rEg.P29T + B, were prepared based on rEg.P29, and a subcutaneous immunization model was established. The intestinal floras of mice in the different immunization groups were analyzed by 16 S rRNA gene sequencing.ResultsThe intestinal microbiota analysis at both immunization time points revealed that Firmicutes, Bacteroidota, and Verrucomicrobiota were the predominant flora at the phylum level, while at the genus level, Akkermansia, unclassified_Muribaculaceae, Lachnospiraceae_NK4A136_group, and uncultured_rumen bacterium were the dominant genera. Some probiotics in the intestines of mice were significantly increased after immunization with the peptide vaccines, such as Lactobacillus_taiwanensis, Lactobacillus_reuteri, Lachnospiraceae_NK4A136_group, Bacteroides_acidifaciens, and so forth. Meanwhile, some harmful or conditionally pathogenic bacteria were decreased, such as Turicibacter sanguinis, Desulfovibrio_fairfieldensis, Clostridium_sp, and so forth, most of which are associated with inflammatory or infectious diseases. Kyoto Encyclopaedia of Genes and Genomes enrichment analysis revealed that the differential flora were enriched in multiple metabolic pathways, primarily biological systems, human diseases, metabolism, cellular processes, and environmental information processing.ConclusionIn this study, we comprehensively analyzed and compared changes in the intestinal microbiota of mice immunized with three peptide vaccines as well as their related metabolic pathways, providing a theoretical background for the development of novel vaccines against E. granulosus.

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“…The serology methods, standardized in each laboratory, have variable sensitivity and specificity due to cross-reactions or weak antigen recognition and thus, low predictive values. Serology for differential CE diagnosis and infection follow-up is a research field in progress, with new laboratory methods such as Raman spectroscopy to evaluate serum samples ( Yue et al., 2020 ) and the proposal of new recombinant antigens, initially examined in infected livestock ( Liang et al., 2020 ). However, no recombinant antigen has demonstrated superiority over hydatid fluid extract for CE diagnosis, and serology is not a useful method for infection or treatment follow-up in humans ( Sánchez-Ovejero et al., 2020 ).…”

Section: Introductionmentioning

confidence: 99%

Characterization of the B-Cell Epitopes of Echinococcus granulosus Histones H4 and H2A Recognized by Sera From Patients With Liver Cysts

Maglioco

Agüero

Valacco

et al. 2022

Front. Cell. Infect. Microbiol.

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Cystic echinococcosis (CE) is a zoonotic disease worldwide distributed, caused by the cestode Echinococcus granulosus sensu lato (E. granulosus), with an incidence rate of 50/100,000 person/year and a high prevalence in humans of 5-10%. Serology has variable sensitivity and specificity and low predictive values. Antigens used are from the hydatid fluid and recombinant antigens have not demonstrated superiority over hydatid fluid. A cell line called EGPE was obtained from E. granulosus sensu lato G1 strain from bovine liver. Serum from CE patients recognizes protein extracts from EGPE cells with higher sensitivity than protein extracts from hydatid fluid. In the present study, EGPE cell protein extracts and supernatants from cell colonies were eluted from a protein G affinity column performed with sera from 11 CE patients. LC-MS/MS proteomic analysis of the eluted proteins identified four E. granulosus histones: one histone H4 in the cell extract and supernatant, one histone H2A only in the cell extract, and two histones H2A only in the supernatant. This differential distribution of histones could reflect different parasite viability stages regarding their role in gene transcription and silencing and could interact with host cells. Bioinformatics tools characterized the linear and conformational epitopes involved in antibody recognition. The three-dimensional structure of each histone was obtained by molecular modeling and validated by molecular dynamics simulation and PCR confirmed the presence of the epitopes in the parasite genome. The three histones H2A were very different and had a less conserved sequence than the histone H4. Comparison of the histones of E. granulosus with those of other organisms showed exclusive regions for E. granulosus. Since histones play a role in the host-parasite relationship they could be good candidates to improve the predictive value of serology in CE.

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Preliminary Evaluation of Recombinant EPC1 and TPx for Serological Diagnosis of Animal Cystic Echinococcosis

Cited by 7 publications

References 42 publications

Modelling diagnostics for Echinococcus granulosus surveillance in sheep using Latent Class Analysis: Argentina as a case study

Modelling diagnostics for Echinococcus granulosus surveillance in sheep using Latent Class Analysis: Argentina as a case study

Changes in intestinal flora of mice induced by rEg.P29 epitope peptide vaccines

Characterization of the B-Cell Epitopes of Echinococcus granulosus Histones H4 and H2A Recognized by Sera From Patients With Liver Cysts

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