Circulating antigen of Aspergillus fumigatus in cortisone-treated mice challenged with conidia: Detection by counterimmunoelectrophoresis

White, Leroy

doi:10.1016/0378-1097(77)90035-0

Cited by 3 publications

(3 citation statements)

References 4 publications

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“…The usefulness of fungal antigen detection in serum and in urine has been demonstrated in animal models of invasive infection with A. fumigatus [13,26]. The BF antigen was chosen as a standard in the ELISA inhibition system described here, as similar models reported in the literature indicated that a heat-stable carbohydrate antigen, derived from the cell wall, is likely to be circulating during the course of disseminated aspergillosis [5,7,18,23].…”

Section: Discussionmentioning

confidence: 98%

Evaluation of a test to detect circulatingAspergillus fumigatusantigen in a survey of immunocompromised patients with proven or suspected invasive disease

Wilson¹,

Hearn²,

Mackenzie³

1987

Med Mycol

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An ELISA for the detection and measurement ofAspergillus antigenaemia has been developed and evaluated by examining sera submitted over a 12-month period from immunocompromised patients with a likelihood of invasive aspergillosis. Results from proven cases ofinvasive aspergillosis confirmed at post-mortem and specimens from individuals with suspected disease showed that tests on single serum samples were often negative. Multiple specimens from the same patient greatly increased the frequency of deteclion. Repeated monitoring of sera from a single patient showed wide fluctuations in antigen level, which was considered to be due partly to the medical regimen to which the patient was subject. Control sera from healthy laboratory personnel were consistently negative, but a number of 'at-risk' patients without other evidence of invasive aspergillosis sometimes had low amounts of antigen.Concentrations of Aspergillus antigen of 100 ng ml-~ or higher were considered to be strongly suggestive of fungal invasion.Patients whose host defense mechanisms have been compromised by immunosuppressive drug therapy are particularly susceptible to invasive fungal infection. In such cases cultural methods of detection and serological tests for antibody are often unsatisfactory.Enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA) and agglutination procedures have been reported which detect antigenaemia in patients with candidosis [25,19,2,8] and coccidioidomycosis [22]. Detector antigens that have been used with success in inhibition systems include Candida cell wall mannan [25,19] and cytoplasmic antigen from Candida yeast phase cells. Similar tests have been evaluated as diagnostic aids for aspergillosis. The antigens used as sensitizing agents for detector plates in ELISA inhibition tests include predominantly carbohydrate entities [18], glycoprotein fractions [14] and purified galactomannan [7]. Some of these antigens have also been used in RIA procedures, namely purified galactomannan [7]

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Section: Discussionmentioning

confidence: 98%

Evaluation of a test to detect circulatingAspergillus fumigatusantigen in a survey of immunocompromised patients with proven or suspected invasive disease

Wilson¹,

Hearn²,

Mackenzie³

1987

Med Mycol

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“…The importance of prompt antifungal treatment in patients with aspergillosis has been demonstrated [2] and it is for this reason that increasing interest is being shown in methods for the detection of Aspergillus-related antigens and metabolites. The usefulness of antigen detection has been demonstrated in a number of animal models of A. fumigatus infection with CIE and RIA [4,30,31,45,59,76,79]. The application of RIA to patients with invasive aspergillosis has also been described [751.…”

Section: Antigen Detection In Invasive Aspergillosmentioning

confidence: 98%

Enzyme-linked immunosorbent assay and its application to the serological diagnosis of fungal infection

Richardson

Warnock

1983

Med Mycol

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With the introduction of the enzyme-linked immunosorbent assay (ELISA), a sensitive technique for detection and quantitation of fungal antibodies and antigens became available. Advantages of ELISA over existing serodiagnostic procedures in medical mycology include Ihe use of objective endpoints, the potential for rapid testing of body fluids and that the equipment necessary to preform basic assays can be both simple and cheap. This review outlines the bases and practical details of the different enzyme-immunoassay techniques and then examines the current status of ELISA procedures that have been applied to particular fungal infections in man and animals.Serological tests are useful as adjuncts to other means of diagnosis of serious fungal infection. These tests are based on the detection of the host's humoral immunological response to fungal infection, or on the detection of products of the fungus itself. Their usefulness depends on the reagents and methods used, on an understanding of their limitations and on judicious interpretation of test findings. Tests for precipitins or agglutinins are often used in the diagnosis of fungal infection [42]. They are useful in patients capable of producing normal levels of immunoglobulins, but are less helpful when applied to immunocompromised hosts where the immune response has been impaired. It is here that sensitive quantitative methods for detection of antibodies or antigen offer the greatest promise and it is not surprising that this field has received so much attention and investment of time.lmmunoassays in various forms have come to play a major role in the serodiagnosis of microbial infection [66]. Immunofluorescence has been widely used in the detection of antibodies and antigens in bacterial, viral and fungal infections. It is, however, rather subjective and labour intensive. In contrast, radioimmunoassay (RIA) is objective and can be automated. However, the reagents used often have a short half-life and there are legal restrictions on the use and disposal of isotopes. Enzyme-immunoassay (EIA), in which enzyme-labelled antibodies or antigens are used, is a more recent technique. Like RIA, it is objective and can be automated. Moreover, it is simple to perform, it is without obvious health risks and the reagents employed have a long shelf life which confers a marked economic advantage over RIA. Specific reagents are available for the detection of individual classes of human immunoglobulin. Established EIA procedures are time consuming but the methods are suited to the development of much more rapid tests. These considerations have led to the development of numerous EIAs, many of which are suitable for use in small laboratories without the need for sophisticated instrumentation.

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“…no. 1146, 1989) and experimental animals (1,8,14,15,19,21,24,27,29). However, with the exception of galactomannan (GM), the antigens used in these assays, and as a consequence the antigens that are circulating or excreted during IA, have not been identified or characterized.…”

Section: Laboratory For Parasitology and Mycologymentioning

confidence: 99%

Pitfalls in immunoblot detection of Aspergillus antigens associated with invasive infection

Wijnands¹,

Leusden²,

Puyk³

et al. 1994

J Clin Microbiol

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strains of 12 species or subspecies of Campylobacter, Helicobacter, and Arcobacter were isolated from 7,680 diarrhetic stool samples. Two hundred and fifty-five (13.8%) of these isolates were dependent on H2-enhanced microaerophilic growth conditions, an essential requirement of C. concisus, C. mucosalis, C. curvus, and C. rectus (7). All 255 clinical isolates plus isolate NCTC 12408 were positive for oxidase and nitrate reductase but were negative for catalase, indoxyl acetate, hippurate, and urease. Every isolate produced abundant H2S, detectable in triple sugar iron agar and often completely blackening lead acetate strips. Isolates tolerate 1% glycine but not 3.5% NaCl. These phenotypic characteristics are shared by C. concisus and C. mucosalis (7). C. mucosalis can be differentiated from C. concisus by its ability to grow at 25°C, its cephalothin sensitivity, and its dirty yellow colony color (6). The issue of growth of C. mucosalis at 25°C is controversial (6, 7). Almost all the clinical isolates, plus isolate NCTC 12408, did not grow at 25°C. Fifteen clinical isolates that were cephalothin sensitive (inhibitory zone sizes of up to 40 mm) and that had a dirty yellow colony color were selected. In other words, as defined by phenotypic criteria, these isolates were C. mucosalis. All 15 clinical isolates plus isolate NCTC 12408 were positive for both arylsulfatase and pyrazinamidase, which is characteristic of C. concisus and not C. mucosalis (1). High-stringency DNA-DNA hybridization studies were performed with theseprobes: C. mucosalis NCTC 1 1000T C. concisus NCTC 11485, C. curvus NCTC 11649T, and C. rectus NCTC 11489T. Isolate NCTC 12408 and all 15 clinical isolates reacted strongly with the C. concisus probe and did not react with any other probe. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (2) confirmed that the 16 isolates were C. concisus. The phenotypic description of C. mucosalis isolates was based on only a few isolates (6). Isolate NCTC 12408 was characterized at NCTC by phenotypic, not molecular, methods as "C. mucosalis-like" (5a). As more and more clinical laboratories adopt filtration and H2-enhanced microaerophilic growth conditions, the problems of differentiating C. mucosalis and C. concisus and related species are becoming apparent. Molecular methods must be used for the precise identification of these underdetected pathogens.

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Circulating antigen of Aspergillus fumigatus in cortisone-treated mice challenged with conidia: Detection by counterimmunoelectrophoresis

Cited by 3 publications

References 4 publications

Evaluation of a test to detect circulatingAspergillus fumigatusantigen in a survey of immunocompromised patients with proven or suspected invasive disease

Evaluation of a test to detect circulatingAspergillus fumigatusantigen in a survey of immunocompromised patients with proven or suspected invasive disease

Enzyme-linked immunosorbent assay and its application to the serological diagnosis of fungal infection

Pitfalls in immunoblot detection of Aspergillus antigens associated with invasive infection

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