scholarly journals Comparison on the performance of Leishmania major-like and Leishmania braziliensis braziliensis as antigen for new world leishmaniasis IgG-immunofluorescence test

1991 ◽  
Vol 33 (6) ◽  
pp. 503-508 ◽  
Author(s):  
Maria Carolina Soares Guimarães ◽  
Beatriz J. Celeste ◽  
Edelma María Corrales L. ◽  
Carlos M.F. Antunes

The performance of an antigen of L. major-like promastigotes for the serological diagnosis of mucocutaneous leishmaniasis in the IgG-immunofluorescent test was compared to that of an antigen of L. braziliensis braziliensis. Each antigen was used to test two hundred and twenty-four sera of etiologies such as mucocutaneous leishmaniasis, deep mycoses, toxoplasmosis, malaria, Chagas' disease, visceral leishmaniasis, anti-nuclear factor, schistosomaiasis, rheumatoid factor and normal controls. Agreement between responses to each antigen was high: 77.2% of leishmaniases sera agreed on a positive or a negative result to both antigens and 91.1 % of control sera. Cross reactivity was restricted to Chagas' disease sera, visceral leishmaniasis, anti-nuclear factor and paracoccidiodomycosis. The quantitative response of leishmaniasis and Chagas' disease sera to both antigens was evaluated by a linear regression; although the y-intercept and the slope were different for each antigen, neither was better than the other in the disclosure of anti-Leishmania antibodies. In the case of Chagas' disease sera the L. major-like antigen was better than L. b. braziliensis' to disclose cross-reacting antibodies.

1996 ◽  
Vol 38 (3) ◽  
pp. 177-185 ◽  
Author(s):  
Ana de Cássia Vexenat ◽  
Jaime M. Santana ◽  
Antonio R.L. Teixeira

We have detected antibodies, in the sera of Chagas disease, Kala-azar and Mucocutaneous leishmaniasis patients, that bind multiple antigens shared between the three causative agents. The Chagas disease sera showed 98 to 100% positive results by ELISA when the Leishmania braziliensis and Leishmania chagasi antigens were used, respectively. The Kala-azar sera showed 100% positive results with Trypanosoma cruzi or L. braziliensis antigens by immunofluorescence assays. The antibodies in the sera of Mucocutaneous leishmaniasis patients showed 100% positive results by ELISA assays with T. cruzi or L. chagasi antigens. Furthermore, the direct agglutination of L. chagasi promastigotes showed that 95% of Kala-azar and 35% of Mucocutaneous leishmaniasis sera agglutinated the parasite in dilutions above 1:512. In contrast, 15% of Chagas sera agglutinated the parasite in dilutions 1:16 and below. Western blot analysis showed that the Chagas sera that formed at least 24 bands with the T. cruzi also formed 13 bands with the L. chagasi and 17 bands with the L. braziliensis. The Kala-azar sera that recognized at least 29 bands with the homologous antigen also formed 14 bands with the T. cruzi and 10 bands with the L. braziliensis antigens. Finally, the Mucocutaneous leishmaniasis sera that formed at least 17 bands with the homologous antigen also formed 10 bands with the T. cruzi and four bands with the L. chagasi antigens. These results indicate the presence of common antigenic determinants in several protozoal proteins and, therefore, explain the serologic cross-reactions reported here.


2002 ◽  
Vol 9 (6) ◽  
pp. 1361-1366 ◽  
Author(s):  
Rosângela Barbosa-de-Deus ◽  
Marcos Luíz dos Mares-Guia ◽  
Adriane Zacarias Nunes ◽  
Kátia Morais Costa ◽  
Roberto Gonçalves Junqueira ◽  
...  

ABSTRACT An antigen (LMS) prepared from Leishmania major-like promastigotes was used in an enzyme-linked immunosorbent assay (ELISA) for the diagnosis of human and dog visceral leishmaniasis. The results were compared with those from the indirect immunofluorescent antibody test (IFAT). A total of 1,822 canine sera were tested, including sera from dogs with visceral leishmaniasis, transmissible venereal tumors, ehrlichiosis, rickettsiosis, or Chagas' disease and sera from healthy dogs. The antigen was also tested with 227 samples of human sera, including sera from patients with visceral, cutaneous, or diffuse cutaneous leishmaniasis and from noninfected individuals, as well as sera from patients with Chagas' disease, toxoplasmosis, rickettsiosis, hepatitis B, schistosomiasis, ascaridiasis, malaria, rheumatoid factor, leprosy and rheumatoid factor, tuberculosis, or leprosy. All dogs and all human patients had a clinical and/or serological and/or parasitological diagnosis. For detecting antibodies in sera from dogs with leishmaniasis, the antigen showed a sensitivity of 98%, specificity of 95%, and concordance of 93% and when used for detecting antibodies in human sera presented a sensitivity of 92%, specificity of 100%, and concordance of 92%. Comparison between ELISA and IFAT demonstrated that ELISA using the LMS antigen yielded more reliable results than IFAT. The LMS antigen displayed no cross-reactivity with sera from patients or dogs that had any of the other diseases tested.


1989 ◽  
Vol 84 (3) ◽  
pp. 309-314 ◽  
Author(s):  
M. G. Morgado ◽  
J. Ivo-dos-Santos ◽  
R. T. Pinho ◽  
E. Argüelles ◽  
J. M. Rezende ◽  
...  

Soluble antigens from epimastigotes of Trypanosoma cruzi were analyzed by western blot in terms of their reactivity with sera from patients with Chagas' disease. In addition, sera from patients with visceral (AVL) and tegumentar leishmaniasis (ATL) were also tested in order to identify cross-reactivities with Trypanosoma cruzy antigens. Twenty eight polypeptides with molecular weights ranging from 14 kDa to 113 kDa were identified with sera from Chagas' disease patients. An extensive cross-reactivity was observed when sera from human visceral leishmaniasis were used, while only a slight cross-reaction was observed with sera from tegumentar leishmaniasis. On the other hand, 10 polypeptidesspecifically reacting with sera from Chagas' disease patients were identified. Among them, the antigens with molecular weights of 46 kDa and 25 kDa reacted with all sera teste and may be good candidates for specific immunodiagnosis of Chagas' disease.


1988 ◽  
Vol 30 (6) ◽  
pp. 411-414 ◽  
Author(s):  
Beatriz J. Celeste ◽  
M. Carolina S. Guimarães ◽  
Edelma Maria Corrales L.

Performance indexes of the peroxidase antibody test were compared to that of the fluorescent antibody test. The peroxidase antibody test had a statistically higher sensitivity and negative predictive value and a higher efficiency than the fluorescent antibody test but its specificity and positive predictive value were within the 95% confidence limits for the values found for the fluorescent antibody test. Such differences did not change when Chagas' disease and visceral leishmaniasis sera were included in index calculations. Statistical analysis showed that the two tests have a substantial degree of agreement but the immunofluorescent test had a specificity index and a positive predictive value equal to 100.0% when Chagas' disease and visceral leishmaniasis sera were not included in the calculations of the performance index; in this instance, a positive test result equals a disclosure of the disease attribute due to the inexistence of false positive results. The enzyme/ protein ratio of the peroxidase conjugate, resulting in heavy or light-labeled conjugates may pose technical problems to its use in serology tests.


2011 ◽  
Vol 53 (5) ◽  
pp. 283-289 ◽  
Author(s):  
Flávia Coelho Ribeiro ◽  
Armando de O. Schubach ◽  
Eliame Mouta-Confort ◽  
Tânia M.V. Pacheco ◽  
Maria de Fátima Madeira ◽  
...  

Indirect immunofluorescence is the method recommended for the diagnosis of visceral leishmanisis in dogs, however, the accuracy of this technique is low and its use on a large scale is limited. Since ELISA does not present these limitations, this technique might be an option for the detection of IgG or specific IgG1 and IgG2 subclasses. Canine ehrlichiosis is an important differential diagnosis of American Visceral Leishmaniasis (AVL). The present study compared ELISA using Leishmania chagasi and Leishmania braziliensis antigen for the detection of anti-Leishmania IgG and subclasses in serum samples from 37 dogs naturally infected with L. chagasi (AVL) and in samples from four dogs co-infected with L. braziliensis and L. chagasi (CI). The occurrence of cross-reactivity was investigated in control serum samples of 17 healthy dogs (HC) and 35 infected with Ehrlichia canis (EC). The mean optical density obtained for the detection of IgG was significantly higher when L. chagasi antigen was used, and was also higher in subgroup VLs (symptomatic) compared to subgroup Vla (asymptomatic). The correlation between IgG and IgG1 was low. The present results suggest that IgG ELISA using homologous antigen yields the best results, permitting the diagnosis of asymptomatic L. chagasi infection and the discrimination between cases of AVL and ehrlichiosis in dogs.


1989 ◽  
Vol 31 (4) ◽  
pp. 228-234 ◽  
Author(s):  
Aoi Masuda ◽  
Sueli Fátima do Nascimento ◽  
Carmem Silvia Guerra ◽  
Gláucia da Silva Paranhos ◽  
Antonio Walter Ferreira

The antigenicity of promastigotes of Leishmania braziliensis braziliensis (L. b.braziliensis) treated with 1% sodium desoxycholate in 10 mM Tris-Hcl pH 8.2 was analysed by immunoblot using as probes sera from American cutaneous leishmaniasis (ACL), American visceral leishmaniasis (AVL), schistosomiasis, malaria and Chagas' disease. The ACL sera reacted constantly with a 60 kD band. No reactivity to this protein was observed with sera from the other diseases above mentioned indicating that the 60 kD protein may be used in serodiagnosis for ACL.


2019 ◽  
Vol 57 (8) ◽  
Author(s):  
Ramona Tavares Daltro ◽  
Leonardo Maia Leony ◽  
Natália Erdens Maron Freitas ◽  
Ângelo Antônio Oliveira Silva ◽  
Emily Ferreira Santos ◽  
...  

ABSTRACT Chimeric T. cruzi antigens have been proposed as a diagnostic tool for chronic Chagas disease (CD) in both settings where Chagas disease is endemic and those where it is not endemic. Antibody response varies in accordance to each T. cruzi strain, presenting challenges to the use of antigens lacking demonstrated cross-reactivity with Leishmania spp. Our group expressed four chimeric proteins (IBMP-8.1, IBMP-8.2, IBMP-8.3, and IBMP-8.4) and previously assessed their diagnostic performance to determine cross-reactivity with Leishmania spp. Here, we validated our findings using serum samples from different Brazilian geographic areas reporting endemic Chagas disease, endemic visceral or American cutaneous leishmaniasis (ACL), or both. Overall, 829 serum samples were evaluated using commercial and IBMP enzyme-linked immunosorbent assays. Due to the absence of a reference assay to diagnosis CD, latent class analysis (LCA) was performed through the use of a statistical model. The incidence of cross-reactivity for ACL-positive samples varied from 0.35% (IBMP-8.3) to 0.70% (IBMP-8.1 and IBMP-8.2). Regarding visceral leishmaniasis (VL)-positive samples, the IBMP-8.2 and IBMP-8.3 antigens cross-reacted with six (3.49%) and with only one sample (0.58%), respectively. No cross-reactivity with either ACL or VL was observed for the IBMP-8.4 antigen. Similarly, no cross-reactions were found when VL-positive samples were assayed with IBMP-8.1. The agreement among the results obtained using IBMP antigens ranged from 97.3% for IBMP-8.2 and 99% for IBMP-8.1 and IBMP-8.3 to 100% for IBMP-8.4, demonstrating almost perfect agreement with LCA. Accordingly, in light of the negligible cross-reactivity with both ACL and VL, we suggest the use of IBMP antigens in regions where T. cruzi and Leishmania spp. are coendemic.


2013 ◽  
Vol 20 (12) ◽  
pp. 1792-1798 ◽  
Author(s):  
Henrique Gama Ker ◽  
Wendel Coura-Vital ◽  
Rodrigo Dian de Oliveira Aguiar-Soares ◽  
Bruno Mendes Roatt ◽  
Nádia das Dores Moreira ◽  
...  

ABSTRACTDiagnosing canine visceral leishmaniasis (CVL) is a critical challenge since conventional immunoserological tests still present some deficiencies. The current study evaluated a prototype flow cytometry serology test, using antigens and fluorescent antibodies that had been stored for 1 year at 4°C, on a broad range of serum samples. Noninfected control dogs andLeishmania infantum-infected dogs were tested, and the prototype test showed excellent performance in differentiating these groups with high sensitivity, specificity, positive and negative predictive values, and accuracy (100% in all analyses). When the CVL group was evaluated according to the dogs' clinical status, the prototype test showed outstanding accuracy in all groups with positive serology (asymptomatic II, oligosymptomatic, and symptomatic). However, in dogs which had positive results by PCR-restriction fragment length polymorphism (RFLP) but negative results by conventional serology (asymptomatic I), serological reactivity was not observed. Additionally, sera from 40 dogs immunized with different vaccines (Leishmune, Leish-Tec, or LBSap) did not present serological reactivity in the prototype test. Eighty-eight dogs infected with other pathogens (Trypanosoma cruzi,Leishmania braziliensis,Ehrlichia canis, andBabesia canis) were used to determine cross-reactivity and specificity, and the prototype test performed well, particularly in dogs infected withB. canisandE. canis(100% and 93.3% specificities, respectively). In conclusion, our data reinforce the potential of the prototype test for use as a commercial kit and highlight its outstanding performance even after storage for 1 year at 4°C. Moreover, the prototype test efficiently provided accurate CVL serodiagnosis with an absence of false-positive results in vaccinated dogs and minor cross-reactivity against other canine pathogens.


2013 ◽  
Vol 7 (05) ◽  
pp. 413-416 ◽  
Author(s):  
Abdolvahab Alborzi ◽  
Gholam R Pouladfar ◽  
Abdolkarim Ghadimi Moghadam ◽  
Armin Attar ◽  
Nima Drakhshan ◽  
...  

Mucocutaneous leishmaniasis, which mostly occurs in the New World, is mainly associated with Leishmania braziliensis and to a lesser degree L. panamensis and L. amazonensis infections. Primary mucosal leishmaniasis is very rare in Iran in spite of high prevalence of cutaneous and visceral leishmanisis. A nine-year-old boy had cutaneous leishmaniaisis for five years involving the left side of his face; he then developed swelling and ulceration of the lip and left side buccal mucosa five months before hospital admission. He had severe swelling of the lower lip and there was ulceration and bleeding of the buccal mucosa. Direct smear revealed leishman bodies and nested PCR confirmed the presence of kinetoplast DNA of L. major in the oral mucosal specimen. The patient received amphotericin B deoxycholate 1 mg/kg/day for one month. The lip and face inflammatory reaction disappeared to nearly normal after one month of therapy. The patient was discharged with ketoconazole (5mg/kg/day) for six weeks. To our knowledge, this is the first report of mucocutaneous leishmaniasis caused by L. major in Iran.


1985 ◽  
Vol 80 (1) ◽  
pp. 17-22 ◽  
Author(s):  
Sergio G. Coutinho ◽  
Marise P. Nunes ◽  
Mauro C. A. Marzochi ◽  
Neide Tramontano

There are areas in the periphery of Rio de Janeiro city where human cases of Visceral and/or Cutaneous Leishmaniasis occur. The parasites have been identified as Leishmania donovani and Leishmania braziliensis braziliensis respectively. A survey for Leishmaniasis was done among 1,342 dogs from those areas using an indirect immunofluorescent test. From the dogs, 616 came from areas where only human cases of Visceral Leishmaniasis occurred, 373 from an area where all human cases were of Cutaneous Leishmaniasis and 353 from a third area (Campo Grande) where both visceral and cutaneous human cases were detected. The prevalence of parasite antibody titers among dogs from areas of Cutaneous Leishmaniasis was significantly higher than that of Visceral Leishmaniasis (8.6% vs. 4.3%, p < 0.02). The highest prevalence was observed among dogs from the area where both diseases are present (12.7%).


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