Estandarización de la técnica Dot-ELISA para la detección de anticuerpos anti-Trypanosoma cruzi y su comparación con ELISA y Western blot

Cervantes-Landín, Alejandra Yunuen; Martínez-Martínez, Ignacio; Reyes, Pedro A.; Shabib, Muslim; Espinoza-Gutiérrez, Bertha

doi:10.1016/j.eimc.2013.05.006

Cited by 7 publications

(10 citation statements)

References 17 publications

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“…For example, the assay has been used to detect human leptospirosis and mycobacteria antigens in patients with pulmonary tuberculosis 31,32. Furthermore, Dot-ELISA has been shown as a quick and reliable assay for the detection of other infectious diseases caused by bacteria, viruses, and parasites such as Chagas, syphilis, Toxocariasis canis, Neospora caninum , chronic Schistosomiasis japonica , cystic echinococcosis, malaria, and neurocysticercosis 33–38…”

Section: Discussionmentioning

confidence: 99%

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Development, optimization, and validation of an in-house Dot-ELISA rapid test based on SAG1 and GRA7 proteins for serological detection of Toxoplasma gondii infections

Teimouri¹,

Modarressi²,

Shojaee³

et al. 2019

IDR

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BackgroundThe aim of the present study was to develop a simple, portable, and rapid assay for serodiagnosis of toxoplasmosis based on recombinant Toxoplasma gondii (T. gondii) SAG1 (rSAG1) and GRA7 (rGRA7) proteins.MethodsThe rSAG1 and rGRA7 proteins were expressed in Escherichia coli (E. coli) and purified in a single step by immobilized metal ion affinity chromatography. The immunoreactivity of the recombinant antigens was tested in an in-house IgG and IgM Dot enzyme-linked immunosorbent assay (Dot-ELISA) for potential use in serodiagnosis of T. gondii infection.ResultsResults from the comparison of in-house rSAG1-Dot-ELISA with ELISA for the detection of anti-Toxoplasma IgG and IgM include sensitivity of 83.7% and 81.2%, specificity of 90.2% and 89.3%, positive predictive values of 85.9% and 68.4%, and negative predictive values of 88.6% and 94.3%, respectively. Sensitivity of 66.2%, specificity of 81.2%, positive predictive values of 71.6%, and negative predictive values of 77.1% were concluded from in-house IgG rGRA7-Dot-ELISA. The sensitivity and specificity of IgM rGRA7-Dot-ELISA included 87.5% and 83.9%, respectively. Sensitivity and specificity of in-house Dot-ELISA for a combination of rSAG1 and rGRA7 included 87.5% and 91.1% for IgG and IgM, respectively. Sensitivity and specificity of a combination of rSAG1 and rGRA7 for the detection of IgM in suspected sera to acute toxoplasmosis were higher than those for the detection of IgG in sera with chronic infections (90.6% and 92% instead of 86.2% and 91.6%, respectively).ConclusionThe highlighted parameters of combined recombinant proteins were more significant than those of single recombinant proteins in in-house Dot-ELISA. These data suggest that the in-house Dot-ELISA based on rSAG1 and rGRA7 combination is a promising diagnostic tool with a similar sensitivity to the native antigens of T. gondii, which can be used for the serodiagnosis of toxoplasmosis in fields as well as less equipped laboratories.

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Section: Discussionmentioning

confidence: 99%

“…Stability of antigens in NC strips has been assessed in other studies. The NC membranes with antigens can be stored for up to 5 months at room temperature without any reactivity changes 38…”

Section: Discussionmentioning

confidence: 99%

Development, optimization, and validation of an in-house Dot-ELISA rapid test based on SAG1 and GRA7 proteins for serological detection of Toxoplasma gondii infections

Teimouri¹,

Modarressi²,

Shojaee³

et al. 2019

IDR

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“…In the present study, we also verified coherence in the interpretation of results between observers (intra and inter-observer). Comparison of DE methodology with different serologic assays has been thoroughly carried out by other authors [ 16 , 39 , 40 ]. It is unanimous that DE reaction is an excellent tool, primarily for seroepidemiological investigations, that is extremely adaptable in extra-laboratory settings and allows the evaluation of a high number of samples while using smaller amounts of antigen.…”

Section: Discussionmentioning

confidence: 99%

Standardization and validation of Dot-ELISA assay for Paracoccidioides brasiliensis antibody detection

Kamikawa

Mendes

Vicentini

2017

J Venom Anim Toxins Incl Trop Dis

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BackgroundParacoccidioidomycosis (PCM) is a neglected systemic mycosis caused by a dimorphic fungus of the Paracoccidioides genus. The standard diagnosis is based on isolation of the fungi in culture, and by microscopic visualization of characteristic multiple budding yeast cells in biological samples. However, in some situations, access to the site of injury prevents the collection of biological material. A variety of immuno-serological techniques has proven useful for allowing inferring diagnosis with a certain degree of certainty, thus optimizing time. The aim of this study was to standardize and validate the Dot-ELISA (DE) assay, comparing it with the serological standard, double immunodiffusion (DI).MethodsIn order to standardize the DE assay, 143 serum samples were used. Out of those, 23 were from apparently healthy patients, 77 were from patients with confirmed PCM and 43 were from patients with other lung infections (tuberculosis, aspergillosis and histoplasmosis). To validate the DE technique, 300 serum samples from patients with PCM clinical suspicion (probable and possible cases) were employed, and these results were compared with those of DI.ResultsThe DE assay showed sensitivity of 91%, specificity of 95.4%, positive predictive value of 96%, negative predictive value of 98.2%, accuracy of 93%, and great precision (k = 0.93). In addition, the nitrocellulose membranes have proved to be viable for using at least 90 days after P. brasiliensis B-339 antigen sensitization.ConclusionDot-ELISA method was found to be an extremely promising tool as serologic screening technique, because of its high sensitivity. Furthermore, Dot-ELISA shows the prospect of being transferred to laboratories of mycoserology including those with fewer resources or even to be used directly in the field. It has an excellent shelf life – membranes coated with antigen can be used for testing without changes in the pattern of reactivity among laboratories – and presents reliable values of sensitivity, specificity, predictive values, accuracy and a high correlation with the serological standard methodology. Based on the present findings, it possible to state that this technique constitutes a remarkable option to be used in routine diagnosis for public health centers.

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“…The 43 selected studies were published between 2010 and 2019, with between 3 and 6 studies being published per year, except for 2015, a year in which there were no publications. 27 articles [9,21,22,24,25,[27][28][29][30][31][32][33][34][35]38,41,42,[44][45][46]48,51,53,55,[57][58][59] evaluated ELISA tests; 14 used RDT [10,23,26,32,37,39,40,43,47,50,52,54,60,61] and 2 [36,56] evaluated both tests (Table 1).…”

Section: Study Featuresmentioning

confidence: 99%

“…According to the nature of the sample, 76.7 % of the investigations were carried out with serum [9,21,22,24-30-36,39,41,42,44,45,48-51,53-60], 7% with venous blood [40,46,61], 7% with venous blood and serum [23,43,52], 4.7% with capillary blood [10,37], 2.3% with serum, plasma, venous blood and capillary [47]; and 2.3 % did not report the type of sample used [39].…”

Section: Study Featuresmentioning

confidence: 99%

Rapid diagnostic tests and ELISA for diagnosing chronic Chagas disease: Systematic revision and meta-analysis

Carrero

Tadger²,

Cuellar³

et al. 2022

PLoS Negl Trop Dis

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Objective To determine the diagnostic validity of the enzyme-linked immunosorbent assay (ELISA) and Rapid Diagnostic Tests (RDT) among individuals with suspected chronic Chagas Disease (CD). Methodology A search was made for studies with ELISA and RDT assays validity estimates as eligibility criteria, published between 2010 and 2020 on PubMed, Web of Science, Scopus, and LILACS. This way, we extracted the data and assessed the risk of bias and applicability of the studies using the QUADAS-2 tool. The bivariate random effects model was also used to estimate the overall sensitivity and specificity through forest-plots, ROC space, and we visually assessed the heterogeneity between studies. Meta-regressions were made using subgroup analysis. We used Deeks’ test to assess the risk of publication bias. Results 43 studies were included; 27 assessed ELISA tests; 14 assessed RDTs; and 2 assessed ELISA and RDTs, against different reference standards. 51.2 % of them used a non-comparative observational design, and 46.5 % a comparative clinical design (“case-control” type). High risk of bias was detected for patient screening and reference standard. The ELISA tests had a sensitivity of 99% (95% CI: 98–99) and a specificity of 98% (95% CI: 97–99); whereas the Rapid Diagnostic Tests (RDT) had values of 95% (95% CI: 94–97) and 97% (95% CI: 96–98), respectively. Deeks’ test showed asymmetry on the ELISA assays. Conclusions ELISA and RDT tests have high validity for diagnosing chronic Chagas disease. The analysis of these two types of evidence in this systematic review and meta-analysis constitutes an input for their use. The limitations included the difficulty in extracting data due to the lack of information in the articles, and the comparative clinical-type design of some studies.

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Estandarización de la técnica Dot-ELISA para la detección de anticuerpos anti-Trypanosoma cruzi y su comparación con ELISA y Western blot

Cited by 7 publications

References 17 publications

<p>Development, optimization, and validation of an in-house Dot-ELISA rapid test based on SAG1 and GRA7 proteins for serological detection of <em>Toxoplasma gondii</em> infections</p>

<p>Development, optimization, and validation of an in-house Dot-ELISA rapid test based on SAG1 and GRA7 proteins for serological detection of <em>Toxoplasma gondii</em> infections</p>

Standardization and validation of Dot-ELISA assay for Paracoccidioides brasiliensis antibody detection

Rapid diagnostic tests and ELISA for diagnosing chronic Chagas disease: Systematic revision and meta-analysis

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