Paracoccidioidomycosis (PCM) is an important systemic fungal disease, particularly among individuals living and working in rural areas of endemicity in Latin America, who, without antifungal therapy, may develop fatal acute or chronic infection. For such patients, the detection of antibody responses by immunodiffusion is of limited value due to false-negative results. In contrast, the detection of Paracoccidioides brasiliensis gp43 circulating antigen may represent a more practical approach to the rapid diagnosis of the disease. Accordingly, an inhibition enzyme-linked immunosorbent assay (inh-ELISA) was developed for the detection of a 43-kDa P. brasiliensis-specific epitope incorporating a species-specific murine monoclonal antibody. With sera from patients with acute and chronic forms of the disease (n ‫؍‬ 81), the overall sensitivity of the test was found to be 95.1%, while specificity was found to be 97.5% compared to that with normal human sera from blood donors

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Abstract. Paracoccidioidomycosis (PCM), the most important human systemic mycosis in Latin America, is known to be caused by at least four different phylogenetic lineages within the Paracoccidioides brasiliensis complex, including S1, PS2, PS3, and Pb01-like group. Herein, we describe two cases of PCM in patients native from the Amazon region. The disease was originally thought to have been caused by P. brasiliensis. Despite the severity of the cases, sera from the patients were negative in immunodiffusion tests using the standard exoantigen from P. brasiliensis B-339. However, a positive response was recorded with an autologous preparation of Paracoccidioides lutzii exoantigen. A phylogenetic approach based on the gp43 and ARF loci revealed high similarity between our clinical isolates and the Pb01-like group. The occurrence of PCM caused by P. lutzii in the Brazilian Amazon (Pará State) was thus proven. The incidence of PCM caused by P. lutzii may be underestimated in northern Brazil.

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IntroductionDisseminated histoplasmosis, a disease that often resembles and is mistaken for tuberculosis, is a major cause of death in patients with advanced HIV disease. Histoplasma antigen detection tests are an important addition to the diagnostic arsenal for patients with advanced HIV disease and should be considered for inclusion on the World Health Organization Essential Diagnostics List.ObjectiveOur objective was to systematically review the literature to evaluate the diagnostic accuracy of Histoplasma antigen tests in the context of advanced HIV disease, with a focus on low- and middle-income countries.MethodsA systematic review of the published literature extracted data on comparator groups, type of histoplasmosis, HIV status, performance results, patient numbers, whether patients were consecutively enrolled or if the study used biobank samples. PubMed, Scopus, Lilacs and Scielo databases were searched for published articles between 1981 and 2018. There was no language restriction.ResultsOf 1327 screened abstracts we included a total of 16 studies in humans for further analysis. Most studies included used a heterogeneousgroup of patients, often without HIV or mixing HIV and non HIV patients, with disseminated or non-disseminated forms of histoplasmosis. Six studies did not systematically use mycologically confirmed cases as a gold standard but compared antigen detection tests against another antigen detection test. Patient numbers were generally small (19–65) in individual studies and, in most (7/10), no confidence intervals were given. The post test probability of a positive or negative test were good suggesting that this non invasive diagnostic tool would be very useful for HIV care givers at the level of reference hospitals or hospitals with the infrastructure to perform ELISA tests. The first results evaluating point of care antigen detection tests using a lateral flow assay were promising with high sensitivity and specificity.ConclusionsAntigen detection tests are promising tools to improve detection of and ultimately reduce the burden of histoplasmosis mortality in patients with advanced HIV disease.

The aim of this study was to isolate and identify Candida species from the oral cavity of denture wearers with denture-related stomatitis who were attended at the University Federal of Pará (Belém City, Pará State, Brazil). A total of 36 denture wearers with denture-related stomatitis were included, and type I (50%), type II (33%) and type III (17%) stomatitis were observed. Candida spp. were isolated from 89% of the cases and included five different Candida species. C. albicans was the most frequently recovered species (78% of the cases), followed by C. famata and C. tropicalis. We observed a significant association between Candida species isolation and unsatisfactory denture condition (p = 0.0017). Our results demonstrated the highly frequency of Candida species isolation in denture wearers with denture-related stomatitis and showed the relationship between these species and poor denture maintenance.

In an attempt to improve the specificity of an enzyme-linked immunosorbent assay (ELISA) for the diagnosis of paracoccidioidomycosis (PCM), sera from patients with PCM were tested using various approaches, such as sodium metaperiodate antigen (gp43) treatment, a serum absorption process with Candida albicans or Histoplasma capsulatum antigens, and dilution of serum in galactose, the main common epitope among pathogenic fungi. The maximum specificity found in this ELISA was 84%. All of these procedures proved inefficient for eliminating all cross-reacting antibodies and obtaining an ELISA specific for PCM diagnosis.Paracoccidioidomycosis (PCM) is a mycotic disease caused by the dimorphic fungus Paracoccidioides brasiliensis and causes a deep mycosis, resulting in a severe chronic granulomatous infection of the skin, mucous membranes, lymph nodes, and internal organs. Almost all South and Central America countries have large regions where PCM is endemic. The definitive diagnosis of PCM includes direct observation of the characteristic multiple-budding cells in biological fluids and tissue sections or isolation of the fungus from clinical materials. For cases in which P. brasiliensis is not observed through direct examination, several serological tests have been used to detect antibodies against the fungus in order to establish the diagnosis (2). gp43, a 43,000-Da glycoprotein from P. brasiliensis, is a major diagnostic antigen in serological assays (1,3,15,18,23,24). It has been demonstrated that gp43 is as good as crude exoantigen preparations (concentrated, dialyzed culture supernatants) (3, 4) for detecting precipitating antibody in immunodiffusion tests, and purified gp43 has also been adapted to several other serological tests, such as immunoprecipitation of 125 I-labeled gp43 (IPP), conventional enzymelinked immunosorbent assay (ELISA), capture enzyme immunoassay (EIA), and passive hemagglutination (5,20,23). Cross reactivity in serological assays from patients with histoplasmosis depends on the test used and seems to involve a carbohydrate epitope. While IPP and capture EIA using gp43 appear specific for PCM diagnosis, experiments were performed to determine the nature of the cross reactivity in sera from those patients with histoplasmosis that reacted with gp43 in the IPP tests. Treatment of gp43 with 0.2 M D-galactose inhibited such cross reactivity, and no cross reactivity was observed with the deglycosylated form of gp43, with EH38, or with the 38,000-Da protein which had been treated with periodate (18). Owing to the N-linked high-mannose nature of the gp43 glycan, deglycosylation was efficiently carried out with endo-␤-N-acetylglucosaminidase H. Since sera from patients with histoplasmosis generally reacted with the high-molecular-weight glycocomplex present in the crude antigen of P. brasiliensis, this reactivity and that with gp43 likely involved recognition of terminal ␤-galactofuranosyl units (24).Travassos et al. (24) related that in general, specific reactions with gp43 are obtained when the ant...

Paracoccidioidomycosis (PCM) is a systemic fungal disease that is particularly important among individuals living and working in rural areas of endemicity in Latin America. Detection of anti-Paracoccidioides brasiliensis antibodies is of limited value due to false-negative results. Detection of P. brasiliensis-gp43 circulating antigen is a practical approach for a specific diagnosis of the disease. In a previous study we described an inhibition enzyme-linked immunosorbent assay able to detect the 43-kDa P. brasiliensis antigen in sera of 100% of patients with the acute form of PCM and in 95.31 and 100% of patients with the chronic multifocal and unifocal forms of PCM. To investigate its potential application for the follow-up of PCM patients during treatment, antigen levels were monitored at regular intervals for up 8 to 12 months in serum samples from 23 patients. The results showed that treatment with itraconazole resulted in decreasing levels of circulating gp43 that were correlated with the reduction of anti-gp43 antibodies. It was also observed that by the end of 12 months of treatment gp43 levels were <5 g/ml in all patients.