A microplate enzyme-immunoassay for toxoplasma antibody.

Voller, A.; Bidwell, D. E.; Bartlett, A.; Fleck, D. G.; Perkins, Margaret; Oladehin, B

doi:10.1136/jcp.29.2.150

Cited by 208 publications

(110 citation statements)

References 6 publications

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“…Immunoenzymatic assay -An in-house immunoenzymatic assay (ELISA) (Voller et al 1976) was performed with the delipidised antigen specific for T. cruzi epimastigotes and processed using Maxisorp plates, as described by Maekelt (1960) and Díaz-Bello et al (2008). ELISAs involving anti-human IgM and IgG alkaline phosphatase conjugates (Sigma Chemical Company, St. Louis, MO, USA) were carried out simultaneously in all sera from individuals at risk.…”

Section: Subjects Materials and Methodsmentioning

confidence: 99%

“…This antigen was validated under a national initiative by the Minister of Health (González et al 2005) and has been used in our laboratory for five decades. Quality control was performed to confirm the reactivity among different lots of antigen.Immunoenzymatic assay -An in-house immunoenzymatic assay (ELISA) (Voller et al 1976) was performed with the delipidised antigen specific for T. cruzi epimastigotes and processed using Maxisorp plates, as described by Maekelt (1960) Laemmli (1970), as modified by Noya et al (1995) and adapted for ChD diagnosis in our laboratory by Zavala-Jaspe et al (2005). Serum detection of five or more specific molecules of the epimastigote antigen was considered a positive result.…”

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confidence: 99%

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The performance of laboratory tests in the management of a large outbreak of orally transmitted Chagas disease

Noya

Díaz-Bello

Colmenares

et al. 2012

Mem. Inst. Oswaldo Cruz

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Orally transmitted Chagas disease (ChD), which is a well-known entity in the Brazilian Amazon Region, was first documented in Venezuela in December 2007, when In Venezuela, effective control measures successfully reduced the annual incidence of Trypanosoma cruzi infections from 10/1,000 individuals in the 1950s to 1/1,000 in the 1980s (Feliciangeli et al. 2003). Since then, the recorded outbreaks of acute Chagas disease (ChD) have been as follows: between 1988-1996, 59 cases occurred in the western states of Venezuela (Añez et al. 1999), after which nine cases involving two deaths occurred during the period of January 2006-March 2007 in the same endemic area (Añez et al. 2007). In 2004 and 2006, the deaths of two children near Caracas were documented ). Nevertheless, an outbreak of orally transmitted ChD (OChD) in a school in a middle class neighbourhood in Caracas was the warning signal that ChD was re-emerging via a different mode of transmission, i.e., the oral route (Alarcón de Noya et al. 2010b Noya et al. 2010b), an outbreak occurred in an urban school that was presumably linked to the ingestion of contaminated home-made juice that had been prepared in a different part of the city where triatomines were present.The aim of this study is to provide information about the parasitological, immunological and molecular laboratory tests used in diagnosing ChD to gleam further knowledge from the largest OChD outbreak ever reported. SUBJECTS, MATERIALS AND METHODSStudy population -The study population consisted of students, teachers and administrative personnel from a school located in the municipality of Chacao in Caracas, Venezuela. Once the primary case was diagnosed (Martín et al. 2009) and the epidemiological link to another case was established (Alarcón de Noya et al. 2010c), parasitological examinations and serological tests were carried out in these cases, in addition to other symptomatic patients who voluntarily requested medical attention at the Tropical Medicine Institute (IMT), Universidad Central de Venezuela in Caracas. After the death of a five-year-old student, most of the school community was managed as part of an on-site emergency, both in the school and at the hospital, where several of the patients were hospitalised. T. cruzi infection was diagnosed by collecting only one tube of blood for serological testing, except for in people who were symptomatic at the time of the interview and from whom additional blood was taken for parasitological or molecular testing (Supplementary data). For logistical and ethical reasons, a unique protocol could not be followed and, consequently, the results are simply presented as they were obtained over time. It was not possible to re-sample the blood of individuals with positive serological tests to isolate the parasite before treatment because the treatment had to begin before the group dispersed for the Christmas holiday.Direct parasitological tests -Observations of the parasite were performed using fresh blood smears as well as Giemsa-stained preparations (W...

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Section: Subjects Materials and Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

The performance of laboratory tests in the management of a large outbreak of orally transmitted Chagas disease

Noya

Díaz-Bello

Colmenares

et al. 2012

Mem. Inst. Oswaldo Cruz

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“…In addition, the need for y-emitting isotopes subjects the users of RIA to a radiation hazard. Enzyme immunoassays (also known as enzyme-linked immunosorbent assays or ELISA) have been developed in an attempt to overcome those problems (3)(4)(5). ELISA is similar in design to solid-phase RIA except that an enzyme is used as the immunoglobulin marker instead of a 'y-emitting isotope.…”

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confidence: 99%

Ultrasensitive enzymatic radioimmunoassay: application to detection of cholera toxin and rotavirus.

Harris¹,

Yolken²,

Krokan³

et al. 1979

Proc. Natl. Acad. Sci. U.S.A.

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Rotavirus and enterotoxin-producing bacteria are major causes of diarrheal disease in humans. A method of rapid diagnosis, ultrasensitive enzymatic radioimmunoassay, has been developed to quantitatively detect cholera toxin and rotavirus. The method uses features of both enzyme-linked immunosorbent assay and radioimmunoassay; however, the sensitivity of the assay is 100-to 1000-fold more sensitive than the two parent assays. Ultrasensitive enzymatic radioimmunoassay should also be useful in measuring other biologically important agents such as drugs and hormones.In recent years, the capability to detect small quantities of biologically important molecules has been greatly expanded by the development of radioimmunoassays (RIA) (1, 2). These assays have been especially useful for the detection of circulating substances, such as hormones, drugs, and infectious agents. However, RIA have certain practical limitations. The short half-life of the isotopes used limits the shelf life of the reagents. In addition, the need for y-emitting isotopes subjects the users of RIA to a radiation hazard. Enzyme immunoassays (also known as enzyme-linked immunosorbent assays or ELISA) have been developed in an attempt to overcome those problems (3-5). ELISA is similar in design to solid-phase RIA except that an enzyme is used as the immunoglobulin marker instead of a 'y-emitting isotope. This enzyme-antibody conjugate is bound to the solid phase by a series of antibody-antigen reactions and essentially converts the substrate to products with a visible yellow color, which can be measured spectrophotometrically. The fact that a single molecule of enzyme is capable of reacting with a large number of substrate molecules provides for amplification and, thus, a high degree of sensitivity (3, 6). Because stable enzymes such as alkaline phosphatase [orthophosphoric-monoester phosphohydrolase (alkaline optimum), EC 3.1.3.1] can be used, ELISA also has the advantage of using reagents with a long shelf life.In practice, however, the sensitivity of ELISA systems has not significantly exceeded that of RIA (7). In order to combine the advantages of both RIA and ELISA, we developed a practical ultrasensitive enzymatic radioimmunoassay (USERIA) for the detection of antigens such as human rotavinus (an important cause of infantile gastroenteritis) and cholera toxin. These assay systems are a 1000-fold more sensitive than both RIA and ELISA for the detection of these agents. MATERIALS AND METHODSReagents.[3H]AMP (generally 3H-labeled, 15 Ci/mmol, New England Nuclear; 1 Ci = 3.7 X 10O°becquerels) was purified by column chromatography with DEAE-Sephadex (A-25; Pharmacia) in a stepwise manner as follows: (i)

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“…Application of ELISA in the diagnosis of toxoplasmosis has been established since 1976 [23]. Since developed up to now, it has been one of the most common biochemical techniques used in research and clinical laboratories, including detection of anti T. gondii antibodies [8,24].…”

Section: Enzyme-linked Immunosorbent Assay (Elisa)mentioning

confidence: 99%

Effective Diagnostic Marker for Serodiagnosis of Toxoplasma gondii Infection: New Developments and Perspectives

Mohamed¹,

Hajissa²

2017

Toxoplasmosis

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Toxoplasmosis is a prevalent parasitic infection caused by an obligate intracellular parasite Toxoplasma gondii. Various methods have been established in the laboratory diagnosis of toxoplasmosis. Among these methods, serological tests are common and provide satisfactory results. However, producing reliable reagents and standard antigen remains difficult and expensive. Replacing native antigens in all current diagnostic kits with standard and reliable reagents are speculated to achieve more sensitive and specific detection that can significantly improve the assay performance. This review provides updated data on toxoplasmosis serodiagnosis. It focuses on the recent trends of producing reliable and standard antigens that have been used in the serological tests of toxoplasmosis, as well as the future direction in this field.

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A microplate enzyme-immunoassay for toxoplasma antibody.

Cited by 208 publications

References 6 publications

The performance of laboratory tests in the management of a large outbreak of orally transmitted Chagas disease

The performance of laboratory tests in the management of a large outbreak of orally transmitted Chagas disease

Ultrasensitive enzymatic radioimmunoassay: application to detection of cholera toxin and rotavirus.

Effective Diagnostic Marker for Serodiagnosis of Toxoplasma gondii Infection: New Developments and Perspectives

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