Evaluation of the western blot in the confirmatory serologic diagnosis of Chagas' disease.

Reiche, Edna Maria Vissoci; Cavazzana, Marina; Okamura, Hirofumi; Tagata, E C; Jankevicius, Shiduca Itow; Jankevicius, José Vítor

doi:10.4269/ajtmh.1998.59.750

Cited by 23 publications

(23 citation statements)

References 22 publications

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“…Although some slight differences in antigen molecular weights were found when our method was compared with other studies that shared the same source of antigens (crude epimastigote antigens) and similar electrophoretic conditions, 12,24,25 our results confirm a similar pattern of reactivity in Chagasic sera.…”

Section: Discussionsupporting

confidence: 76%

“…12,[17][18][19][20][21][22][23] The main limitation of this technique is that it is difficult to perform interlaboratory comparisons, mainly due to the heterogeneity of the strains and antigenic preparations used. In addition, the size of the slab gels and the concentration of polyacrylamide used as well as the different molecular weight markers all introduce additional variations in band separation and the establishment of respective molecular weights.…”

Section: Discussionmentioning

confidence: 99%

“…This variability between different methods and laboratories is probably caused by the use of different strains of T. cruzi, different antigenic fractions, and different procedures, the immunogenetic capacity of the parasite strain, and the variation in humoral response that each individual develops during infection. 12 Furthermore, in patients who are coinfected with other pathogens, especially Leishmania, the possibility of crossreactivity frequently limits the specificity of serological results. This is considered a major drawback for the serological diagnosis of Chagas disease in areas endemic for both T. cruzi and Leishmania spp.…”

Section: Introductionmentioning

confidence: 99%

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Identification of a Western Blot Pattern for the Specific Diagnosis of Trypanosoma cruzi Infection in Human Sera

Riera

Vergés²,

Iniesta³

et al. 2012

The American Journal of Tropical Medicine and Hygiene

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Abstract. A Western blot (WB) method using a lysate from Trypanosoma cruzi (Maracay strain) epimastigotes was evaluated. Serum samples from 37 patients with confirmed Chagas disease (cohort I), 27 Spanish patients with visceral leishmaniasis caused by Leishmania infantum (cohort II), and 28 Colombian patients with cutaneous leishmaniasis caused by L. panamensis and negative serology for Chagas disease (cohort III) were tested. The negative controls were 55 healthy seronegative subjects for T. cruzi and Leishmania; 28 of the negative controls were from a region endemic for Chagas disease and Leishmania (cohort IV), and 27 of the negative controls were from a non-endemic area for Leishmania and T. cruzi (cohort V). A homogeneous standard band pattern consisting of six antigenic bands corresponding to 28, 32, 38, 39, 40, and 48 kDa was recognized simultaneously for all Chagasic patients' sera. Sera from Leishmania-infected patients showed a heterogeneous band pattern that was easily differentiated from the pattern of patients with Chagas disease. WB with T. cruzi epimastigote antigen is an efficient method for diagnosis and may be used as an alternative to confirm T. cruzi and detect cross-reactivity with Leishmania.

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Section: Discussionsupporting

confidence: 76%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Identification of a Western Blot Pattern for the Specific Diagnosis of Trypanosoma cruzi Infection in Human Sera

Riera

Vergés²,

Iniesta³

et al. 2012

The American Journal of Tropical Medicine and Hygiene

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“…Two other important findings were that proteins with higher molecular masses (Ͼ118 kDa) reacted with negative control sera and that proteins other than those of 15, 21 to 22, 63, 74 to 75, 110, and 118 kDa reacted with the antibodies in the sera of very few patients with CL and VL. However, Reiche et al (27) reported that the detection of proteins of 18, 30, 39, 45, 52, 64, 70, and 82 kDa in CL patient sera is not enough to establish a positive result in Brazilian CL patients.…”

Section: Discussionmentioning

confidence: 99%

Serodiagnosis of Anthroponotic Cutaneous Leishmaniasis (ACL) Caused byLeishmania tropicain Sanliurfa Province, Turkey, Where ACL Is Highly Endemic

Zeyrek

Korkmaz

Özbel

2007

Clin Vaccine Immunol

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In this study, we aimed to evaluate the validity of the conventional enzyme-linked immunosorbent assay (ELISA) and the Western blotting test for the diagnosis of anthroponotic cutaneous leishmaniasis (ACL) using serum samples obtained from 51 patients with parasitologically proven nontreated CL (NonT-CL patients) and 62 patients under treatment for CL (UT-CL patients). Additionally, 29 serum samples obtained from patients with parasitologically and serologically proven visceral leishmaniasis (VL) were also used as positive controls, and serum samples from 43 blood donors were used as negative controls. All sera were diluted to the same dilution (1/100). Leishmania infantum MON-1 was used as the antigen in the conventional ELISA. The sera of 27 (93.1%) of 29 VL patients were seropositive by ELISA, while the sera of 40 (78.4%) of 51 NonT-CL patients and 43 (69.3%) of 62 UT-CL patients were seropositive by the conventional ELISA. The absorbance values of the CL patients' sera were significantly lower than the absorbance values of the VL patients' sera. Bands between 15 and 118 kDa were detected in two groups of CL patients. Among all bands, the 63-kDa band was found to be more sensitive (88.5%). When we evaluated the Western blotting results for the presence of at least one of the diagnostic antigenic bands, the sensitivity was calculated to be 99.1%. By using serological tests, a measurable antibody response was detected in most of the CL patients in Sanliurfa, Turkey. It is also noted that this response can be changed according to the sizes, types, and numbers of lesions that the patient has. The Western blot test was found to be more sensitive and valid than the conventional ELISA for the serodiagnosis of ACL. In some instances, when it is very difficult to demonstrate the presence of parasites in the smears, immunodiagnosis can be a valuable alternative for the diagnosis of ACL.

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“…41,42 A serum sample was considered positive when it recognized at least one antigenic band from a group of three with the highest frequency of recognition. The result was considered negative when the serum sample showed no reactivity with these diagnostic antigenic bands.…”

Section: Methodsmentioning

confidence: 99%

Evaluation of antigens from various Leishmania species in a Western blot for diagnosis of American tegumentary leishmaniasis.

Gonçalves¹,

Reiche²,

Filho³

et al. 2002

The American Journal of Tropical Medicine and Hygiene

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Abstract. A Western blot method that uses antigens from culture promastigote forms of Leishmania (Viannia) braziliensis, L. (Leishmania) amazonensis, L. (Leishmania) tropica, and a trypanosomatid (strain 268T) isolated from naturally infected tomatoes was evaluated for laboratory diagnosis of American tegumentary leishmaniasis (ATL). Serum samples were obtained from 108 patients with ATL (group I), 23 chagasic patients (group II), 32 patients with other diseases (group III), and 78 healthy individuals (group IV). The overall analysis showed a sensitivity of 76.90%, 90.40%, 78.50%, and 87.90%, a specificity of 100%, 93.80%, 87.80%, and 77.10%, a positive predictive value of 100%, 94.00%, 89.50%, and 72.50%, a negative predictive value of 75.70%, 90.00%, 75.40%, and 90.20%, and a concordance coefficient kappa of 0.7358, 0.8400, 0.6491, and 0.6287 for L.(L.) tropica, and strain 268T antigens, respectively. The antigenic profile recognized by serum samples from patients with ATL and with Chagas' disease permits serologic distinction between these infections.

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Evaluation of the western blot in the confirmatory serologic diagnosis of Chagas' disease.

Cited by 23 publications

References 22 publications

Identification of a Western Blot Pattern for the Specific Diagnosis of Trypanosoma cruzi Infection in Human Sera

Identification of a Western Blot Pattern for the Specific Diagnosis of Trypanosoma cruzi Infection in Human Sera

Serodiagnosis of Anthroponotic Cutaneous Leishmaniasis (ACL) Caused byLeishmania tropicain Sanliurfa Province, Turkey, Where ACL Is Highly Endemic

Evaluation of antigens from various Leishmania species in a Western blot for diagnosis of American tegumentary leishmaniasis.

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