Standardization of the Dot Enzyme-Linked Immunosorbent Assay (Dot-ELISA) for Human Visceral Leishmaniasis

Pappas, Michael G.; Hajkowski, Ruta; Hockmeyer, Wayne T.

doi:10.4269/ajtmh.1984.33.1105

Cited by 36 publications

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“…Dot-ELISA test, a modification of the standard ELISA test, offers a simple and less expensive tool for toxocariasis detection. The dot-ELISA has been successfully adapted for the detection of parasitic diseases in humans as leishmaniasis, schistosomiasis, toxoplasmosis, and hydatidosis (Pappas et al 1984, Boctor et al 1987, Rogan et al 1991, Elsaid et al 1995. A dot-ELISA for diagnosis of human toxocariasis was described (Camargo et al 1992).…”

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Evaluation of the dot enzyme-linked immunosorbent assay in comparison with standard ELISA for the immunodiagnosis of human toxocariasis

Roldán

Cornejo

Espinoza

2006

Mem. Inst. Oswaldo Cruz

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A dot enzyme-linked immunosorbent assay (dot-ELISA) was standardized using excretory-secretory antigens ofKey words: Toxocara canis -toxocariasis -dot enzyme-linked immunosorbent assay -ELISA Human toxocariasis is a world-wide helminthic zoonosis due to the human infection by larvae of Toxocara canis, the common ascarid of dogs, and also by the cat ascarid T. cati (Schantz & Glickman 1983, Despommier 2003. The incidence of human toxocariasis is unknown because toxocariasis is not a communicable disease in the majority of the countries. However, many cases of this disease have been reported throughout the world (Glickman & Schantz 1981, Despommier 2003.Humans are infected by ingestion of embryonated T. canis eggs. Children playing in areas contaminated with dog faeces are in higher risk, because of their likelihood of ingesting soil. Prevalence of Toxocara infestation of dogs and the resulting contamination of the ground is relatively high in many countries all over the world. Reported data range from 0 to 93% for dog infestation (Glickman & Schantz 1981) and 15 to 78% for soil contamination (Gillespie 1988, Magnaval et al. 2001. It has been determined that in Peru from 24 to 80% of soil samples in public playgrounds and parks are contaminated with Toxocara eggs (Buitrón 1976, Guerrero 1995, Lescano et al. 1998, Chavez et al. 2000, Dávalos et al. 2000.Although the clinical features vary, two syndromes are recognized: visceral larva migrans (VLM) and ocular larva migrans (OLM). VLM is usually detected in young children (1 to 5 years of age) with a history of geophagia and/or exposure to puppies. It is a self-limited, rarely lethal disease characterized by fever, cough, wheeze, pallor, malaise, asthma, weight loss, hepatomegaly, and marked eosinophilia (Schantz 1989 mus and, more rarely, eye pain (Shields 1984). Another clinical manifestations of Toxocara infection are "common toxocariasis" in adults and "covert toxocariasis" in children (Schantz 1989, Magnaval et al. 2001.Diagnosis of toxocariasis is based on clinical and serological data because of the difficulty in detecting larvae from tissues. The test currently employed for the serodiagnosis of toxocariasis is ELISA using excretory-secretory antigens from T. canis second-stage larvae (TES) (De Savigny et al. 1979, Jacquier et al. 1991. However, this technique has some drawbacks, including the need for trained personnel, requirement of special equipment, the lack of reproducible reading due to plate-to-plate variation, and may be troublesome to perform under field conditions. Dot-ELISA test, a modification of the standard ELISA test, offers a simple and less expensive tool for toxocariasis detection. The dot-ELISA has been successfully adapted for the detection of parasitic diseases in humans as leishmaniasis, schistosomiasis, toxoplasmosis, and hydatidosis (Pappas et al. 1984, Boctor et al. 1987, Rogan et al. 1991, Elsaid et al. 1995. A dot-ELISA for diagnosis of human toxocariasis was described (Camargo et al. 1992). The present study was conducted to standa...

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confidence: 99%

Evaluation of the dot enzyme-linked immunosorbent assay in comparison with standard ELISA for the immunodiagnosis of human toxocariasis

Roldán

Cornejo

Espinoza

2006

Mem. Inst. Oswaldo Cruz

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“…However, dot-ELISA appears to be one of the more sensitive and least affinity-dependent procedures. 3 In contrast with antibody response against serotype B polysaccharide, which has been shown to be mediated mostly by IgM, the response against the OMCs antigens using dot-ELISA seems to be mediated by other isotypes, such as IgG and IgA. This assay was also useful in the longitudinal analysis of the antigen-specific immunoglobulin titers for use in immunological studies of this severe disease.…”

Section: Discussionmentioning

confidence: 99%

“…2 Dot-ELISA has been used to detect a variety of bacterial and protozoal antigens. [3][4][5][6] Coll et al, 1998, developed a dot-ELISA that uses monoclonal antibodies for detection of serogroup B meningococcal antigens in cerebrospinal fluid that has become a valuable tool in the detection of meningococci. Furthermore, in 1989 Oprandy & Sippel 7 used a monoclonal antibody that was immobilized on a nitrocellulose membrane for an antigen capture assay.…”

Section: Introductionmentioning

confidence: 99%

Comparison of dot-ELISA and standard ELISA for detection of Neisseria meningitidis outer membrane complex-specific antibodies

Belo¹,

Farhat²,

Gaspari³

2010

Braz J Infect Dis

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Dot-ELISA using the outer membrane complex antigens of Neisseria meningitidis as a target was standardized for rapid detection of meningococcal-specific antibodies in human serum. We investigated the level of meningococcal-specific IgG, IgA, and IgM in serum using dot-ELISA with outer membrane antigens prepared from Neisseria meningitidis serotype B:4.19:P1.15,3,7,9 (a strain isolated from a Brazilian epidemic). The dot-ELISA is based on the same principles as the standard ELISA and is useful for detection of anti-N. meningitidis B antibodies in serum of patients with meningococcal infections. For the assay, outer membrane complexes (OMCs) were absorbed by nitrocellulose membrane and blocked with a 5% skim milk solution. Serum samples were drawn upon hospital admission and during convalescence from patients with meningococcal septicemia, and single samples were drawn from uninfected controls. We retrospectively examined a total of 57 serum samples: 35 from patients infected with N. meningitidis B, 12 from patients infected with Haemophilus influenzae b, and 10 from health individuals. When performed at room temperature, dot-ELISA took approximately four hours to perform, and the optimum antigen concentration was 0.42 μg per dot. The specificity of IgG, IgM, and IgA demonstrates that dot-ELISA using OMCs from N. meningitidis B as a target is suitable for serologic verification of clinically suspected meningococcal disease in patients and for titer determination of antibodies produced during different phases of natural infection. Furthermore, the sensitivity of dot-ELISA was comparable to that of standard ELISA. Overall, dot-ELISA is simple to perform, rapid, and low cost. Further validation of the test as a screening tool is required.

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“…Other serum samples included were taken from 37 dogs with other confirmed infectious diseases; i.e., ehrlichiosis (18), distemper (15) coronavirus and parvovirus (4), from other endemic leishmaniasis areas in Catalonia. Eighty sera from dogs from nonendemic leishmaniasis areas were also included: Tenerife Island (Canary Island, Spain) (26), Switzerland (19) and Sweden (35). Lymph node aspirates from 107 dogs from the Priorat focus were examined by Giemsa staining and cultured in NNN (Novy-McNeal-Nicolle) biphasic blood agar medium.…”

Section: Methodsmentioning

confidence: 99%

“…19 Modifications introduced in this assay, like the use of a polyvalent conjugate as protein A and the use of a bio-dot apparatus to simplify the manipulation, resulted in a simple, highly specific, and sensitive test. The test has long been used in our laboratory in different seroepidemiologic studies on dogs from endemic leishmaniasis areas, from nonendemic areas and from veterinary clinics.…”

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Serologic Diagnosis of Canine Leishmaniasis by Dot-ELISA

et al. 1997

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Abstract.A dot-enzyme-linked immunosorbent assay (ELISA) using protein A-peroxidase was evaluated as a diagnostic test for canine leishmaniasis. The test results were in agreement with parasitologic diagnosis and indirect immunofluorescence assay results. The sensitivity of the test calculated on 31 dogs with positive parasitologic examination was 90% when a titer of 1/800 was established as a cutoff and 100% when a titer of 1/400 was established. The specificity calculated on the canine population from nonendemic areas was 100% when both titers were established. Nevertheless, in endemic areas titers near the cutoff need careful interpretation. The results of this study demonstrate that dot-ELISA protein A using a bio-dot apparatus is highly suitable for seroepidemiologic field work.Canine leishmaniasis is endemic throughout the Mediterranean basin. The disease is caused by the same parasite as that causing human leishmaniasis, Leishmania infantum. 17 The domestic dog and the European fox (Vulpes vulpes) are incriminated as being the main reservoir of the parasite. 1,16,22 The clinical features of canine leishmaniasis vary widely. The 3 major symptoms are depilation, onycogryphosis, and emaciation, and their observation in an endemic area strongly suggests leishmaniasis.3 Nevertheless, because of the variability of the symptoms and the high frequency of asymptomatic or oligosymptomatic dogs in endemic areas, 2,10 the clinical diagnosis of the disease is difficult.A definitive diagnosis of leishmaniasis is established by direct observation of the intracellular stage of the parasite in bone marrow, lymph node aspirates, or skin lesions. The conventional parasitologic methods of diagnosis have low sensitivity; 2,15 their usefulness is limited by the number of parasites in the target organs, contamination of cultures, and the ability of strains to grow in vitro. For effective screening of infected dogs, it is necessary to adopt a simple, sensitive, and specific diagnostic test.Indirect immunofluorescence assay (IFA) is one of the most commonly used immunologic tests for the detection of L. infantum infections in humans and in the dog reservoir, 2,14 but its application requires a high level of skill and experience and large laboratory facilities.In 1983, a dot enzyme-linked immunosorbent assay (ELISA) for the serologic diagnosis of visceral human Received for publication April 12, 1996. leishmaniasis was described as a rapid and economical method that was as sensitive, specific, and reproducible as the more time-consuming ELISA procedure. 18 The test has been used 4,8,11,12 in the diagnosis of human visceral leishmaniasis in the Old World and the diagnosis of cutaneous and mucocutaneous human leishmaniasis and canine visceral leishmaniasis in the New World.In this paper, we report a similar dot-ELISA for the serodiagnosis of canine leishmaniasis following a previously described procedure.19 Modifications introduced in this assay, like the use of a polyvalent conjugate as protein A and the use of a bio-dot apparat...

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Standardization of the Dot Enzyme-Linked Immunosorbent Assay (Dot-ELISA) for Human Visceral Leishmaniasis

Cited by 36 publications

References 0 publications

Evaluation of the dot enzyme-linked immunosorbent assay in comparison with standard ELISA for the immunodiagnosis of human toxocariasis

Evaluation of the dot enzyme-linked immunosorbent assay in comparison with standard ELISA for the immunodiagnosis of human toxocariasis

Comparison of dot-ELISA and standard ELISA for detection of Neisseria meningitidis outer membrane complex-specific antibodies

Serologic Diagnosis of Canine Leishmaniasis by Dot-ELISA

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