Detection of Thyroid-Stimulating Immunoglobulins by Use of Enzyme-Fragment Complementation

Sandrock, Tanya; Terry, Alan H.; Martin, Jeff C.; Erdogan, Evrim; Meikle, Wayne

doi:10.1373/clinchem.2007.100396

Cited by 1 publication

(2 citation statements)

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“…These assays are capable of differentiating stimulatory from blocking and neutral antibodies and their use in clinical laboratory practice has the potential to predict relapsed patients or Graves' disease remission more accurately [10]. In this report, we looked to improve upon the efficiency of an early version of the cell-based bioassay [11]. We hypothesized that some genetic alterations to the TSHR expression cassette would aid accumulation of cAMP, and thus sensitivity.…”

Section: Introductionmentioning

confidence: 99%

“…bs_bs_banner Clinical and Experimental Immunology ORIGINAL ARTICLE doi:10.1111/j. 1365-2249.2012.04648.x In this report, we looked to improve upon the efficiency of an early version of the cell-based bioassay [11]. We hypothesized that some genetic alterations to the TSHR expression cassette would aid accumulation of cAMP, and thus sensitivity.…”

Section: Introductionmentioning

confidence: 99%

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Measurement of thyroid stimulating immunoglobulins using a novel thyroid stimulating hormone receptor–guanine nucleotide-binding protein, (GNAS) fusion bioassay

Pierce

Sandrock

Gillespie

et al. 2012

Clinical and Experimental Immunology

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SummaryHyperthyroidism, defined by overproduction of thyroid hormones, has a 2-3% prevalence in the population. The most common form of hyperthyroidism is Graves' disease. A diagnostic biomarker for Graves' disease is the presence of immunoglobulins which bind to, and stimulate, the thyroid stimulating hormone receptor (TSHR), a G-protein coupled receptor (GPCR). We hypothesized that the ectopically expressed TSHR gene in a thyroid stimulating immunoglobulin (TSI) assay could be engineered to increase the accumulation of the GPCR pathway second messenger, cyclic AMP (cAMP), the molecule measured in the assay as a marker for pathway activation. An ectopically expressing TSHR-mutant guanine nucleotidebinding protein, (GNAS) Chinese hamster ovary (CHO) cell clone was constructed using standard molecular biology techniques. After incubation of the new clone with sera containing various levels of TSI, GPCR pathway activation was then quantified by measuring cAMP accumulation in the clone. The clone, together with a NaCl-free cell assay buffer containing 5% polyethylene glycol (PEG)6000, was tested against 56 Graves' patients, 27 toxic thyroid nodule patients and 119 normal patients. Using receiver operating characteristic analysis, when comparing normal with Graves' sera, the assay yielded a sensitivity of 93%, a specificity of 99% and an efficiency of 98%. Total complex precision (within-run, across runs and across days), presented as a percentage coefficient of variation, was found to be 7·8, 8·7 and 7·6% for low, medium and high TSI responding serum, respectively. We conclude that the performance of the new TSI assay provides sensitive detection of TSI, allowing for accurate, early detection of Graves' disease.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%