Development and validation of an improved enzyme-linked immunosorbent assay for the detection of thyroglobulin autoantibodies in canine serum samples

Iversen, Lars; Jensen, A. L.; Høier, R.; Skydsgaard, Mikala; Kristensen, Frans Brandt

doi:10.1016/s0739-7240(98)00040-x

Cited by 21 publications

(24 citation statements)

References 25 publications

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“…In the assay used by Haines et al [16], the number of TgAA positive dogs in non-hypothyroid dogs was high, so that some of the positive dogs may have been false-positive. These "false-positive" results were explained by the fact that the epitope recognized by TgAA in dogs with hypothyroidism differs from that in normal dogs positive for TgAA [17]. Thus, our hypothesis holds that there is the pathogenic epitopes recognized by TgAA on cTg in hypothyroid dogs.…”

mentioning

confidence: 78%

“…Purification of canine thyroglobulin: cTg was purified according to the methods described in the reports [17,30,33]. Blood samples were collected from two clinically normal, purebred beagle dogs.…”

Section: Methodsmentioning

confidence: 99%

“…Detection of autoantibody specific for thyroglobulin (Tg) is an indispensable laboratory subject for the diagnosis of Hashimoto's thyroiditis in human medicine. The antibody specific for Tg has been also found in a high proportion of clinically diagnosed hypothyroid dogs [2,14,16,17]. Recently the reliability of enzyme-linked immunosorbent assay (ELISA) techniques for Tg autoantibody (TgAA) detection in dogs was compared with that of thyroid biopsy, and was demonstrated as excellent diagnostic methods with high sensitivity and specificity [1,17,24].…”

mentioning

confidence: 99%

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Recognition Pattern of Thyroglobulin Autoantibody from Hypothyroid Dogs to Tryptic Peptides of Canine Thyroglobulin

Tani

Shimizu

Sasai

et al. 2003

The Journal of Veterinary Medical Science

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ABSTRACT. Circulating thyroglobulin autoantibody (TgAA) was analyzed using the Western immunoblot for determination of the dominant epitopes recognized by TgAA on tryptic peptides of canine thyroglobulin (cTg) in hypothyroid dogs. TgAA was measured in hypothyroid dogs, non-hypothyroid dogs with skin diseases and clinically normal dogs. Five of the 7 hypothyroid dogs, 1 of the 8 dogs with skin diseases and 1 of the 4 normal dogs were positive for TgAA. Four of the 5 TgAA-positive hypothyroid dogs were Golden Retri evers, and 3 of them showed high antibody titers. The sera of TgAA positive-dogs reacted to several peptides, and their patterns varied from sample to sample. Sera from 3 dogs with high titers of TgAA reacted broadly to high molecular weight peptides ranging from 45 to 90 kDa. These Western immunoblot patterns of the sera were disappeared after pretreatment with sufficient amount of intact cTg. All serum samples of both TgAA positive dogs and negative controls reacted to low molecular weight peptides ranging from 15 to 20 kDa. These immunoblot patterns of the sera were not disappeared even after pretreatment with sufficient amount of intact cTg. These findings show the possibility that the epitopes recognized by TgAA depend upon individual dogs with hypothyroidism and these aut oantibodies recognize conformational epitopes on the cTg molecule.

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mentioning

confidence: 78%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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Recognition Pattern of Thyroglobulin Autoantibody from Hypothyroid Dogs to Tryptic Peptides of Canine Thyroglobulin

Tani

Shimizu

Sasai

et al. 2003

The Journal of Veterinary Medical Science

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“…ELISA for thyroglobulin autoantibody: Canine TgAA in the serum samples was detected by the method of Iversen et al [13], with minor modifications [22]. Horseradish peroxidase-conjugated sheep anti-dog IgG (Serotec Ltd, Kidlington, Oxford, UK) was used.…”

Section: Animalsmentioning

confidence: 99%

“…Thyroglobulin-specific antibodies have also been found in a high proportion of clinically diagnosed hypothyroid dogs [2,9,11,13]. Thyroperoxidase antibodies were not found in one study [23], but uncharacterized microsomal antibodies have been reported in others [10,15,25].…”

mentioning

confidence: 99%

Proliferative Responses to Canine Thyroglobulin of Peripheral Blood Mononuclear Cells from Hypothyroid Dogs

Tani

Nabetani

Sasai

et al. 2005

The Journal of Veterinary Medical Science

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ABSTRACT. The immune responses of hypothyroid dogs to canine thyroglobulin (cTg) were evaluated for the proliferative ability of peripheral blood mononuclear cells (PBMC). PBMC from three hypothyroid dogs with high titers of thyroglobulin autoantibody (TgAA) and 3 clinically normal dogs were cultured with 5, 10, or 20 µg/ml of cTg for 72 hr. The proliferative responses of the cells were determined by the level of incorporated BrdU. The numbers of cells expressing Thy-1, CD4, CD8 and IgG in the PBMC were counted by the immunofluorescence method. Proliferative responses to cTg were observed in the cells from hypothyroid dogs. The number of cells expressing IgG and CD8 in the hypothyroid dogs tended to be high compared with the clinically normal dogs. The CD4 + cells in cultures from hypothyroid dogs increased depending upon the amount of cTg. There was a significant (P<0.05) positive correlation between the number of CD4 + cells and the concentration of cTg in the cultures from hypothyroid dogs. These findings suggest a possible relationship between canine hypothyroidism and cellular immunity. Loss of self tolerance to thyroid antigens in CD4 + T cells may play an important role in the development of canine hypothyroidism.

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Standardization and validation of an alkaline phosphatase-linked immunoassay to quantify a plant-derived antibody directed against the hepatitis B virus surface antigen

Leyva

Hernández

González

et al. 2004

Biochemical and Biophysical Research Communications

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Development and validation of an improved enzyme-linked immunosorbent assay for the detection of thyroglobulin autoantibodies in canine serum samples

Cited by 21 publications

References 25 publications

Recognition Pattern of Thyroglobulin Autoantibody from Hypothyroid Dogs to Tryptic Peptides of Canine Thyroglobulin

Recognition Pattern of Thyroglobulin Autoantibody from Hypothyroid Dogs to Tryptic Peptides of Canine Thyroglobulin

Proliferative Responses to Canine Thyroglobulin of Peripheral Blood Mononuclear Cells from Hypothyroid Dogs

Standardization and validation of an alkaline phosphatase-linked immunoassay to quantify a plant-derived antibody directed against the hepatitis B virus surface antigen

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