Analysis of Paracoccidioides brasiliensis exoantigens stability

Silva, Décio Fragata da

doi:10.1590/s1678-91992006000400012

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“…This finding has major advantages for public health laboratories, since obtaining the culture filtrate is significantly less costly and requires less complex procedures than those required to obtain purified and/or recombinant antigens. Another point that deserves special attention is the good performance (in liters) and the excellent stability of the culture filtrate, more than 15 years after production [ 37 ]. Furthermore, we found that the use of culture filtrate also provides good index of sensitivity, specificity, accuracy, positive and negative predictive values, secured the intermediate repeatability and reproducibility of the Dot-ELISA assay.…”

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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Section: Discussionmentioning

confidence: 99%