Evaluation of native 8kDa antigen based three immunoassays for diagnosis of cystic echinococcosis in sheep

Jeyathilakan, N.; Basith, S. Abdul; John, Lalitha; Chandran, N. D. J.; Raj, G. Dhinakar; Churchill, R. R.

doi:10.1016/j.smallrumres.2013.10.012

Cited by 10 publications

(10 citation statements)

References 39 publications

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“…Currently, data are limited on recombinant diagnostic antigens for detection of E. granulosus infection in sheep [ 11 , 12 ], and the diagnostic sensitivity of the recombinant proteins used was very low in these reports. Many methods, such as the flow through technique, enzyme linked immuno electrotransfer blot [ 13 , 14 ], enzyme-linked immunosorbent assay (ELISA) [ 15 ], counter-immunoelectrophoresis, and the latex agglutination test [ 14 ] have been developed for the diagnosis of CE in sheep. However, these assays use natural E. granulosus hydatid cyst fluid antigens, which are difficult and expensive to prepare, and cannot be commercialized.…”

Section: Introductionmentioning

confidence: 99%

Molecular characterization and serodiagnostic potential of a novel dithiol glutaredoxin 1 from Echinococcus granulosus

Song

Yan

et al. 2016

Parasites Vectors

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BackgroundThe larval stage of Echinococcus granulosus is the etiological agent of cystic echinococcosis (CE), which causes serious morbidity and mortality in many areas. There is no reliable method to monitor sheep CE. Here, we characterize E. granulosus glutaredoxin 1 (Eg-Grx1) and report an improved immunodiagnostic method for CE.MethodsWe cloned and expressed recombinant Eg-Grx1 and generated antibodies. We analyzed the location of the protein in different parasite stages by fluorescence immunohistochemistry, detected the immunogenicity of recombinant Eg-Grx1, and developed an indirect ELISA (iELISA) for CE serodiagnosis.ResultsEg-Grx1 is a classic dithiol Grx with several GSH-binding motifs. Native Eg-Grx1 protein was distributed in the tegument of protoscoleces, the whole germinal layer, and the parenchymatous tissue of adult worms. Recombinant Eg-Grx1 exhibited good immunoreactivity to CE-infected sheep serum. An iELISA using this antigen showed specificity of 64.3 % (9/14) and sensitivity of 1:3200, and the diagnostic accordance rate was 97.9 % (47/48) compared with the results of necropsy.ConclusionWe characterized a novel Grx (Eg-Grx1) from a parasitic helminth and present a comprehensive analysis of the sequence and structure of this protein. The recombinant Eg-Grx1 protein showed good potential serodiagnostic performance, and we established an iELISA method, which may contribute to the surveillance of sheep CE in epidemic areas.

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Section: Introductionmentioning

confidence: 99%

Molecular characterization and serodiagnostic potential of a novel dithiol glutaredoxin 1 from Echinococcus granulosus

Song

Yan

et al. 2016

Parasites Vectors

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“…Cystic echinococcosis is a major zoonosis infecting humans and a wide range of animals, causing global health problems and economic losses worldwide (Sarkar et al, 2017). Diagnosis of CE by means of serology has limited support in clinical practice (Golassa et al, 2011;Jeyathilakan et al, 2014). Due to the poor specificity, complex composition and unstable antigen sources of natural proteins (Tamarozzi et al, 2014), establishing an accurate and sensitive immunologic diagnostic method based on recombinant protein has become crucial for CE detection.…”

Section: Discussionmentioning

confidence: 99%

“…The detection of animal CE mainly relies on necropsy and molecular and serological diagnosis (Tamarozzi et al, 2014;Manzanoromán et al, 2015;Ito et al, 2016). For laboratory detection and surveillance, lab-based serology such as enzyme-linked immunosorbent assays (ELISAs) (Simsek and Koroglu, 2004), counter immunoelectrophoresis (Zhang and Mcmanus, 2013), latex agglutination (Jeyathilakan et al, 2014), and colloidal gold immunochromatographic strips (Zhuo et al, 2017) have been developed. However, because of the very long period of incubation and different organ localization, detection, and surveillance of CE infection is often inaccurate (error rate 15.4 %) (Gottstein, 1992;Gatti et al, 2007;Moro and Schantz, 2009).…”

Section: Introductionmentioning

confidence: 99%

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

Liu

Song

et al. 2020

Front. Cell. Infect. Microbiol.

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Animal cystic echinococcosis (CE) is one of the most important helminthic diseases and affects many mammalian intermediate hosts. Practical and effective diagnosis is crucial for animal CE control. Two different recombinant antigens derived from Echinococcus granulosus, Echinococcus protoscolex calcium binding protein 1 (rEg-EPC1) and thioredoxin peroxidase (rEg-TPx), were evaluated in this study to detect the specific immunoglobulin G (IgG) in sheep and goat with CE by the indirect enzyme-linked immunosorbent assays. The diagnostic effect of the above-listed proteins was determined to their sensitivity and specificity and compared with hydatid cyst fluid, two previously reported immunogenic recombinant proteins (dihydrofolate reductase and P29), and two commercial kits available in China. Of these, the best diagnostic results were obtained in the anti-TPx IgG ELISA, with 92.6% sensitivity, 98.8% specificity, and no cross-reactivity with anti-Eg95 IgG. Recombinant E. granulosus thioredoxin peroxidase shows good potential for serological diagnosis of animal cystic echinococcosis.

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“…Ultrasound and other diagnostic methods have been used to improve the accuracy of diagnosis. However, compared with indirect ELISA, such methods have disadvantages of expense and scarce materials [ 13 , 14 , 16 ]. Moreover, compared with traditional methods, indirect ELISA has the advantages of high reproducibility and stability of antigen source.…”

Section: Discussionmentioning

confidence: 99%

“…Traditional diagnostic methods for animal E. granulosus are based on necropsy in abattoirs [ 10 , 11 ], which commonly use judgment by macroscopic observation and have a high error rate (15.4%) [ 12 ]. There are many other ways to diagnose CE in sheep, including the flow through technique (flow anion exchange chromatography) [ 13 ], the latex agglutination test [ 14 ], and enzyme-linked immunosorbent assay (ELISA) [ 15 , 16 ]. These methods especially using the hydatid cyst fluid as antigens to diagnose CE of sheep are difficult to develop into commercial products.…”

Section: Introductionmentioning

confidence: 99%

Expression, Tissue Localization and Serodiagnostic Potential of Echinococcus granulosus Leucine Aminopeptidase

Yan

et al. 2018

IJMS

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Echinococcus granulosus is the causative agent of cystic echinococcosis (CE), a widespread parasitic zoonosis. Leucine aminopeptidases (LAPs) of the M17 peptidase family have important functions in regulating the balance of catabolism and anabolism, cell maintenance, growth and defense. In this study, we presented a bioinformatic characterization and experimentally determined the tissue distribution characteristics of E. granulosus LAP (Eg-LAP), and explored its potential value for diagnosis of CE in sheep based on indirect ELISA. Through fluorescence immunohistochemistry, we found that Eg-LAP was present in the tegument and hooks of PSCs, the whole germinal layer and adult worm parenchymatous tissue. Western blotting results revealed that the recombinant protein could be identified using E. granulosus-infected sheep serum. The diagnostic value of this recombinant protein was assessed by indirect ELISA, and compared with indirect ELISA based on hydatid fluid antigen. The sensitivity and specificity rEgLAP-ELISA were 95.8% (23/24) and 79.09% (87/110), respectively, while using hydatid fluid as antigen showed the values 41.7% (10/24) and 65.45% (72/110). This is the first report concerning leucine aminopeptidase from E. granulosus, and the results showed that Eg-LAP belong to M17 peptidase families, and that it is involved in important biological function of E. granulosus. Furthermore, rEg-LAP is appropriate for diagnosing and monitoring CE in sheep in field. Development of a rapid test using rEg-LAP to diagnose sheep CE deserves further study.

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Evaluation of native 8kDa antigen based three immunoassays for diagnosis of cystic echinococcosis in sheep

Cited by 10 publications

References 39 publications

Molecular characterization and serodiagnostic potential of a novel dithiol glutaredoxin 1 from Echinococcus granulosus

Molecular characterization and serodiagnostic potential of a novel dithiol glutaredoxin 1 from Echinococcus granulosus

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

Expression, Tissue Localization and Serodiagnostic Potential of Echinococcus granulosus Leucine Aminopeptidase

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