An analysis of antibodies against <i>Aspergillus jumigatus</i> in bovine serum by iminunoblotting and enzyme‐linked immunosorbent assays

Jensen, Henrik Elvang; Latgé, Jean‐Paul

doi:10.1111/j.1699-0463.1995.tb01087.x

Cited by 11 publications

(8 citation statements)

References 22 publications

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“…[2][3][4]14,[18][19][20][21]27,31,37 Because systemic mycoses including aspergillosis are difficult to diagnose clinically, most cases are not diagnosed until histopathologic evaluation is completed as part of a postmortem examination. 2-4,14,18-2l,27,3l,37 Attempts to culture the etiologic agents of systemic mycoses including Aspergillus spp.…”

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confidence: 99%

“…In most of these techniques, polyclonal antibodies have been applied as the primary reagents, 12,13,15,19,21,22,24,26,32,33 whereas monoclonal antibodies (MAbs) have only been utilized for a limited number of fungal species and a few studies. 6,7,16,20,23,34,40 The use of polyclonal antibodies rather than MAbs in immunodiagnostic techniques has some disadvantages, e.g., the time-consuming and labor-intensive heterologous absorption of polyclonal antibodies, which is necessary to achieve sufficient specificity. Moreover, the -production of polyclonal an-tibodies is difficult to standardize because it depends on the antigen used for immunization, the methods used, and the animal selected, its housing, and its nutrition.…”

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Development of Murine Monoclonal Antibodies for the Immunohistochemical Diagnosis of Systemic Bovine Aspergillosis

Jensen¹,

Aalbæk

Lind³

et al. 1996

J VET Diagn Invest

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Abstract. Murine monoclonal antibodies (MAbs) against water-soluble somatic antigens (WSSA) and the wall fraction (WF) from Aspergillus fumigatus were produced by fusion of splenocytes from immunized BALB/c mice with mouse myeloma X63-Ag 8.653 cells. The supernatants of in vitro cultured hybridomas were initially screened for reactivity with the WSSA and the WF from A. fumigatus and WSSA of other fungi in an enzymelinked immunosorbent assay (ELISA). Supernatants reacting only with A. fumigatus antigens were subsequently screened for homologous and heterologous reactivity with immunohistochemical techniques using formalinfixed, paraffin-embedded tissues from experimentally infected mice. Because of a high immunohistochemical reactivity with homologous fungi, 4 MAbs raised against A. fumigatus WSSA and WF were selected for a further evaluation of cross-reactivity (diagnostic specificity) in immunohistochemical and immunoblotting assays. In immunohistochemical assays, all MAbs raised against WSSA cross-reacted heavily with a number of other fungal species. All 4 MAbs (MAb-WF-AF-1-4) raised against the WF reacted strongly with hyphae of Aspergillus spp.; hyphae of Scedosporium apiospermum were also strongly labeled by MAb-WF-AF-3 and-4. The 2 specifically reacting MAbs (MAb-WF-AF-1 and-2) were of the IgM biotype and were precipitating, and in immunoblotting experiments both bound to a 106-kD antigen of the WF, whereas they did not bind to WSSA of A. fumigatus. One of the 2 aspergillosis-specific MAbs (MAb-WF-AF-1) was used to screen 145 mycotic lesions of cattle. The diagnoses on bovine lesions obtained by MAb-WF-AF-1 were compared with results based on reactivity with heterologously absorbed polyclonal antibodies and, for some lesions, to culture results. In the vast majority of lesions (n = 133), the MAb-WF-AF-1 and the polyclonal anti-Aspergillus antibodies reacted in a similar pattern, i.e., positively in 41 aspergillosis lesions and negatively in 92 zygomycotic lesions. Hyphae in 3 of 12 lesions that were not stained by the polyclonal antibodies reacted with the specific MAb-WF-AF-1; i.e., aspergillosis was diagnosed. The characteristics of the 2 MAbs (MAb-WF-AF-1 and-2) raised against the WF of A. fumigatus in ELISA and immunoblotting and immunohistochemical assays justify their application for the in situ diagnosis of systemic aspergillosis of cattle.Aspergillus fuimigatus is the main cause of systemic bovine aspergillosis, a disease occuring worldwide and of great importance in cattle production. [2][3][4]14,[18][19][20][21]27,31,37 Because systemic mycoses including aspergillosis are difficult to diagnose clinically, most cases are not diagnosed until histopathologic evaluation is completed as part of a postmortem examination. 2-4,14,18-2l,27,3l,37 Attempts to culture the etiologic agents of systemic mycoses including Aspergillus spp. may give false results because of several difficulties, e.g., growth failure, overgrowth by contaminants, recovery of a fungus different from that suggested by the morpholo...

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confidence: 99%

Development of Murine Monoclonal Antibodies for the Immunohistochemical Diagnosis of Systemic Bovine Aspergillosis

Jensen¹,

Aalbæk

Lind³

et al. 1996

J VET Diagn Invest

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“…For this reason, we established a cut‐off value of X + 3SD, with serum dilutions as low as 1/500 and 1/3000, in order to differentiate between healthy and affected animals or, at least, eliminate false positives. Precedents can be found in veterinary medicine in a study to determine the level of anti‐ Aspergillus IgG in cases of bovine aspergillosis using an ELISA methodology (Jensen and Latge, 1995). In each of the 41 control animals used, antibodies against A. fumigatus were detected, but in only one of these animals was the level moderately higher than the level that was considered positive.…”

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confidence: 99%

The Value of the Determination of Anti‐AspergillusIgG in the Serodiagnosis of Canine Aspergillosis: Comparison with Galactomannan Detection

García

Caballero

Cruzado

et al. 2001

Journal of Veterinary Medicine, Series B

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Diagnosis of canine aspergillosis is difficult using currently available methods. It often passes unnoticed or is diagnosed in the later phases of the disease. We developed an ELISA technique to detect anti-Aspergillus antibodies in canine serum using an Aspergillus antigenic mycelial extract, which could then be used for the diagnosis of canine aspergillosis. We used a cut-off of X + 3SD obtained from 20 control sera. The test was performed on 46 dogs with lesions indicating possible aspergillosis and gave nine positive results: one systemic mycosis, two discospondylitis, one uveitis, two bronchopulmonary processes and three rhinitis. We compared this methodology with the PLATELIA technique in the follow-up of the affected dogs, obtaining the same limitations as in the diagnosis of human aspergillosis. We consider our ELISA technique using sera samples a speedy, safe and reliable method which enables us to follow up the evolution of the disease and the efficacy of the therapy chosen. A definitive diagnosis must still take into account the results of other tests such as clinical examination, radiographic studies, endoscopy and biopsy.

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“…Despite the potential for an Aspergillus ‐specific diagnosis, it seems that anti‐ Aspergillus antibody quantification may not be useful due to the high background in healthy individuals, which could be caused by the ubiquitous dispersal of Aspergillus in the environment, and hence repeated exposure of birds. Anti‐ Aspergillus antibody detection in dogs, cats, horses, and cattle is also difficult, where healthy controls show positivity, although titers might be higher in infected animals. Overall, the environmental presence of Aspergillus makes antibody detection tests unreliable and must be interpreted with care and in the context of other diagnostic results.…”

Section: Serologymentioning

confidence: 99%

The current status of avian aspergillosis diagnoses: Veterinary practice to novel research avenues

Savelieff¹,

Pappalardo

Azmanis

2018

Veterinary Clinical Pathol

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Aspergillus fungal infections continue to be a significant cause of morbidity and mortality in birds that can, in part, be attributed to the lack of a diagnostic "gold standard" for Aspergillus infection, and which delays the diagnosis, treatment, and outcome of avian patients. At present, none of the available methods in veterinary care can detect aspergillosis early enough and with the accuracy, precision, and specificity required of an ideal diagnostic tool. Therefore, researching methods of Aspergillus detection is still an active area of inquiry, and novel techniques continue to emerge. This review will provide a brief overview of current clinical methods, with an emphasis on avian care, in addition to a series of techniques in development that could offer distinct advantages over existing methods.

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An analysis of antibodies against Aspergillus jumigatus in bovine serum by iminunoblotting and enzyme‐linked immunosorbent assays

Cited by 11 publications

References 22 publications

Development of Murine Monoclonal Antibodies for the Immunohistochemical Diagnosis of Systemic Bovine Aspergillosis

Development of Murine Monoclonal Antibodies for the Immunohistochemical Diagnosis of Systemic Bovine Aspergillosis

The Value of the Determination of Anti‐AspergillusIgG in the Serodiagnosis of Canine Aspergillosis: Comparison with Galactomannan Detection

The current status of avian aspergillosis diagnoses: Veterinary practice to novel research avenues

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