Immunoenzymatic Assay (Elisa) in Mucocutaneous Leishmaniasis, Kala-Azar, and Chagas' Disease: An Epimastigote Trypanosoma cruzi Antigen Able to Distinguish between Anti-Trypanosoma and Anti-Leishmania Antibodies *

Guimarães, Morgana Rodrigues; Celeste, Beatriz Julieta; Castilho, Euclides Ayres de; Mineo, José Roberto; Diniz, J. M. P.

doi:10.4269/ajtmh.1981.30.942

Cited by 41 publications

(21 citation statements)

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“…Thus, the rORFF-based ELISA appears more sensitive than the DAT, which is known for a high degree of specificity and sensitivity for VL. 7,16 The slightly elevated OD 405 values for noninfected individuals from endemic regions compared with those from non-endemic regions may reflect subclinical infection. 24 Since the assay has not yet been fully optimized with respect to antigen amount and serum dilution, it is conceivable that an assay can be developed that will accurately predict the parasitologic status of all VL patients.…”

Section: Resultsmentioning

confidence: 99%

Serodiagnosis of leishmaniasis with recombinant ORFF antigen.

Raj¹,

Ghosh²,

Dole³

et al. 1999

The American Journal of Tropical Medicine and Hygiene

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Abstract. The serodiagnostic potential of recombinant ORFF protein (rORFF) from Leishmania infantum was assessed by ELISA. Of 49 sera from confirmed cases of visceral leishmaniasis (VL), all were seropositive using 5 ng of rORFF and serum diluted 1:20, while only 38 were positive with 500 ng of soluble antigen (SA) and 44 were positive by a direct agglutination test. There was also a positive correlation between spleen size and level of seropositivity with rORFF or SA. The reciprocal endpoint titer with rORFF was 1,280 for sera from VL patients, but Ͻ 20 with sera from malaria, filariasis, and tuberculosis patients, as well as with sera from healthy individuals from endemic and non-endemic areas. Sera from 10 confirmed cutaneous leishmaniasis cases from Turkey were negative or only weakly positive with rORFF, although 9 were positive with SA. Thus, rORFF protein appears useful as a sensitive reagent for the differential diagnosis of VL caused by the Leishmania donovani complex.Protozoan parasites of the genus Leishmania cause a spectrum of human disease ranging from self-limiting cutaneous leishmaniasis (CL) to the lethal visceral leishmaniasis (VL). There is a general correlation between the species of Leishmania and the type of disease, with the Leishmania donovani complex causing VL or kala-azar. Routine diagnosis of VL has long been based on the microscopic detection of parasites in smears of lymph node, bone marrow, or splenic aspirates or culturing the parasite from patient material. These surgical procedures for diagnosis are painful and dangerous 1 ; thus, there is an urgent need for safe and reliable tests for specific diagnosis of leishmaniasis.Serologic tests that have been developed and evaluated clinically include an immunofluorescent antibody test (IFAT), 2-4 an ELISA with either whole parasites or purified antigens, 5-10 a dot-ELISA, 11,12 immunoblot analysis, 13,14 and the direct agglutination test (DAT). [15][16][17][18] The DAT using microtiter plates for the diagnosis of VL and American CL gives satisfactory results and is easy to perform. 19 It is a simple technique with high sensitivity and specificity. 15,18 One of its major drawbacks is that it has no prognostic value for evaluating the progression of the disease. The DAT is used as a routine serologic test for VL in India, and the parameters of the test have been established under local conditions with very little false positivity. 18,20 In this paper, we report the serodiagnostic potential for Indian VL of recombinant ORFF protein (rORFF) that is encoded in the LD1 locus of chromosome 35 of L. infantum. 21 The rORFF appears to be specifically recognized by sera from patients with VL compared with those with CL. Furthermore, the sensitivity of detection appears to be greater than with total L. donovani promastigote soluble antigen, suggesting the utility of rORFF protein as a sensitive and differential diagnostic reagent. MATERIALS AND METHODSPatient sera. Sera were collected from 49 patients with clinically confirmed VL in the Sahibganj ...

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Section: Resultsmentioning

confidence: 99%

Serodiagnosis of leishmaniasis with recombinant ORFF antigen.

Raj¹,

Ghosh²,

Dole³

et al. 1999

The American Journal of Tropical Medicine and Hygiene

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“…The pellets were resuspended in 200 volumes of 0.15 M NaOH containing 0.014 M PMSF (Sigma) followed by three cycles of sonication at 40 Hz for 20 s each time in an ice bath [11]. The suspension was slowly shaken (150 rpm) for 6 h at 4 o C. The pH was adjusted to 7.2, and the material kept under slow shaking overnight.…”

Section: Parasite and Antigensmentioning

confidence: 99%

“…Among the serological tests used for the diagnosis are the direct agglutination, indirect immunofluorescence (IIF), and enzyme immunoassay (EIA). Some EIA techniques for the ACL diagnosis were described by several authors that used L. braziliensis [11][12][13][14] and other Leishmania species [15][16][17] as antigen, and they detected IgG [12,14,15,17,18], IgM [12] or IgA antibodies [13].…”

mentioning

confidence: 99%

Diagnosis of American cutaneous leishmaniasis by enzyme immunoassay in patients from Northern Paraná State, Brazil

Yoneyama¹,

Peder²,

Lonardoni³

et al. 2007

Braz J Infect Dis

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American cutaneous leishmaniasis (ACL) is an endemic disease caused by Leishmania parasites. The ACL diagnosis is commonly accomplished by parasitological and immunological methods. The objective of this work was to evaluate the enzyme immunoassay (EIA) for ACL laboratorial diagnosis. IgG antibodies against Leishmania (Viannia) braziliensis promastigotes were researched. For the method standardization 240 sera were used: 72 from patients with positive parasitological diagnosis, 38 from normal individuals and 113 from individuals with other pathologies. The sensibility was 93% and the specificity was 70%. Regarding diagnosis, EIA showed 97% positivity in patients with ACL and 97% negativity in patients with other cutaneous lesions. The EIA presented a good performance when used for diagnosis, thus, it may become an important tool for it and for further ACL epidemiological studies in endemic areas.

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“…Two antigens were prepared for each parasite species: one followed a preparation 6 , consisting of an alkaline promastigote extract and another consisting of the same alkaline extract to which 1 ul% of phenylmethane sulphonyl fluoride (PMSF) was added prior to cell disruption 16 .…”

Section: Seramentioning

confidence: 99%

“…Sera were drawn from male patients (86.0%) and female (13.8%), 7 to 83 years of age. On previous assays sera had tested positive for anti-Leishmania IgG class antibodies by a L. major-like enzyme-linked immunoassay ELISA 6 thirteen sera were drawn from in-patients at the Department of Dermatology, University of São Paulo Medical School, and 16 were drawn at regional offices of the Fundação Nacional de Saúde, Brazilian Ministry of Health. Twentytwo sera previously found to be negative for a n t i -L e i s h m a n i a a n t i b o d i e s a n d a n t iTrypanosoma cruzi antibodies by IgGimmunofluorescence 11 and IgG-Elisa, were used as negative controls.…”

mentioning

confidence: 99%

Evaluation of Leishmania antigens preparation and storage for use in enzyme immunoassays

Celeste

Guimarães

Souza³

1997

Rev. Soc. Bras. Med. Trop.

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One disadvantage of growing Leishmania (V.) braziliensis in liquid medium is its cost due to the need for addition of fetal calf serum to Schneider's Drosophila medium 12 . The same does not occur with L. major-like promastigotes which grow abundantly in culture media such as LIT (liver infusion tryptose) 4 a quality to be looked for when considering its use on mucocutaneous leishmaniasis seroepidemiology research where large quantities of parasites are needed; also, its use as antigen results in good performance indexes of sensitivity, specificity and positive predictive value on IFI tests, IgG-ELISA and Dot-ELISA 2 8 10 . L. major-like has been widely used for leishmaniasis seroepidemiologic research, mainly for IFI tests 7 14 , but until this moment its stability as antigen for any serology test had yet to be evaluated. The same happens with Leishmania (V.) braziliensis, although its use as antigen for seroepidemiological work has been much less than L. major-like's due to the difficulty for culturing the parasite as it grows poorly on liquid culture media in spite of the use of especially formulated products 1 .The present report investigates the influence of time and temperature on the storage of alkaline extracts of L. major-like and L. (V.) braziliensis promastigotes and also the addition of PMSF by investigating their influence on the GMT of mucocutaneous leishmaniasis standard sera. The study on time and temperature was intended as a means to critically ascertain optimum storage and temperature for the antigens, an important step for serum surveys where the use of only one antigen batch is prefered. As proteases inhibitors 18 have been used in parasitic antigenic extracts in an attempt to minimize the action of proteolitic enzymes we thought interesting to critically evaluate its influence on the alkaline antigen performance by means of its influence on serum GMT. MATERIAL AND METHODS Sera

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Immunoenzymatic Assay (Elisa) in Mucocutaneous Leishmaniasis, Kala-Azar, and Chagas' Disease: An Epimastigote Trypanosoma cruzi Antigen Able to Distinguish between Anti-Trypanosoma and Anti-Leishmania Antibodies *

Cited by 41 publications

References 0 publications

Serodiagnosis of leishmaniasis with recombinant ORFF antigen.

Serodiagnosis of leishmaniasis with recombinant ORFF antigen.

Diagnosis of American cutaneous leishmaniasis by enzyme immunoassay in patients from Northern Paraná State, Brazil

Evaluation of Leishmania antigens preparation and storage for use in enzyme immunoassays

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