Cláudia Regina De Marchi

Observações sobre o TESA blot no diagnóstico sorológico da doença de Chagas

TESA blot was compared with indirect hemagglutination, indirect immunofluorescence and ELISA tests. In sera from 30 participants infected with Trypanosoma cruzi, and in 30 non infected the four techniques produced entirely equivalent results, all positive and all negative, respectively. In cases admitted to be inconclusive or in visceral leishmaniasis, frequent false positives were detected. However, TESA blot contributed with the least proportion of them.

False-positive results of a rapid K39-based strip test and Chagas disease.

BACKGROUND The definitive diagnosis of visceral leishmaniasis (VL) requires invasive procedures with demonstration of amastigotes in tissue or promastigotes in culture. Unfortunately, these approaches require laboratory materials not available in poor countries where the disease is endemic. The correct diagnosis of VL is important, and made more difficult by the fact that several common tropical diseases such as malaria, disseminated tuberculosis, and enteric fever share the same clinical presentation. Serological tests have been developed to replace parasitological diagnosis in the field. A commercially available K39-based strip test for VL has been developed for this purpose. The endemic area of leishmaniasis in Brazil overlaps the endemic area of Chagas disease, a disease that can cause false-positive serological test results. The aim of this study was to evaluate the incidence of false-positive exams using a rapid test for VL in patients with Chagas disease. METHODS A rapid test based on the recombinant K39 antigen of Leishmania was used in: (1) 30 patients with confirmed Chagas disease, (2) 30 patients with a serological diagnosis of Chagas disease by ELISA, indirect immunofluorescence, indirect hemagglutination, and chemiluminescence, (3) 30 healthy patients from a non-endemic area as the control group, (4) 30 patients with confirmed VL, and (5) 20 patients with proved cutaneous leishmaniasis. RESULTS The sensitivity and specificity of the rapid strip test were 100% when compared with healthy volunteers and those with confirmed Chagas disease. One false-positive result occurred in the group with Chagas disease diagnosed by serological tests (specificity of 96%). CONCLUSION The rapid test based on recombinant K39 is a useful diagnostic assay, and a false-positive result rarely occurs in patients with a serological diagnosis of Chagas disease.

Avaliação da resposta imune humoral frente a antígenos de Strongyloides venezuelensis

Strongyloidiasis affects 30 million people in 70 countries. This enteral parasitosis is usually diagnosed using parasitological tests based on hydrotropism or thermotropism of larvae eliminated in feces, but these tests have been shown to have low sensitivity. In this study, antigenic extracts were tested by means of ELISA, immunoblotting and IFI, using filariform larvae of Strongyloides venezuelensis, a parasite of rodents that shows cross-reactions with Strongyloides stercoralis epitopes. Sensitivity of 89, 85 and 57% for the ELISA reaction and 100, 100 and 96% for immunoblotting with the SAL, ZWIP and ZW antigens, and specificity of 90, 60 and 81% for ELISA and 96, 92 and 91% for immunoblotting with the same antigens, were found in these assays.

Trypanosoma cruzi parasitemia observed in immunocompromised patients: the importance of the artificial xenodiagnosis

Trypanosoma cruzi parasitemia observed in immunocompromised patients (transplant or positive HIV) occurred more frequently by the artificial xenodiagnosis method (10/38) compared with hemoculture (2/38), given the same quantity of blood. Other ways of diagnosis, like mice inoculation (5/38), QBC and buffy coat (2/38), were evaluated also. This result showed the importance of the artificial xenodiagnosis. The other techniques increased only one more patient positive.

Comportamento do método quimioluminescente-ELISA em relação a resultados considerados discordantes por meio de três técnicas convencionais para diagnóstico da doença de Chagas

When indirect hemagglutination, indirect immunofluorescence and enzyme-linked immunosorbent assay are used together for serologically diagnosing Chagas disease, results that are considered discordant sometimes occur because there is disagreement between what these tests indicate. The availability of the chemiluminescent ELISA method enabled tests on 200 serum samples that had previously produced discordant results from the three above-mentioned methods. CL-ELISA revealed that 193 of these samples were negative and seven were positive. The use of this new procedure provides further support for understanding this subject, but more concrete advances will depend on documentation with blood analyses from people previously demonstrated to be unquestionably infected or uninfected with Trypanosoma cruzi.

Avaliação da sensibilidade da diluição 1/20 pela reação de imunofluorescência indireta, no diagnóstico sorológico da doença de Chagas

Sera from people in the chronic stage of Chagas disease, whose infection had been parasitologically validated, were assayed by using the indirect immunofluorescence test to evaluated its performance at the 1:20 dilution. All tests were consistently positive at 1:20 and higher dilutions, even in the presence of concomitant infection with the human immunodeficiency virus (HIV). It is thus valid, into the light of this experiment, to take into account the remarkable sensitivity of such serological test at the above mentioned dilution.

Evaluation of the humoral immune response against purified and recombinant mucins from Trypanosoma cruzi

The clinical laboratory diagnosis of chronic Chagas disease is routinely carried out using serological tests which employ crude extracts obtained from the non-infective epimastigote form of Trypanosoma cruzi. The existence in these preparations of numerous antigens and epitopes, which are shared by other microorganisms, in addition to the low serum dilution used in the conventional serological tests, frequently leads to inconclusive or false-positive results. Here, we have developed two new chemiluminescent ELISA tests: the first employs as antigens purified amastigote and trypomastigote mucins, collectively named TAKK-P; and the second uses as an antigen a recombinant mucin, named TSSAII. Both tests were simultaneously evaluated for sensitivity and specificity using panels of sera with positive, negative or inconclusive results for conventional serology of Chagas disease, and also heterologous sera. The CL-ELISA with TAKK-P showed 100% sensitivity and 99.8 specificity, whereas the CL-ELISA with TSSA-II showed 89.9% sensitivity and 95.6% specificity.