Optimisation of an immunohistochemistry method for the determination of androgen receptor expression levels in circulating tumour cells.
BMC Cancer
; 14: 226, 2014 Mar 28.
Article
em En
| MEDLINE
| ID: mdl-24674711
ABSTRACT
BACKGROUND:
AZD3514 inhibits and down regulates the androgen receptor (AR) and has undergone clinical trials in prostate cancer. To provide proof-of-mechanism (POM) in patients, an immunohistochemistry (IHC) method for determination of AR in circulating tumour cells (CTC) was developed and validated.METHODS:
After an assessment of specificity validation focused on intra- and inter-operator reproducibility utilising a novel modification of incurred sample reanalysis (ISR). ß-Content γ-confidence tolerance intervals (BCTI) and Cohen's Kappa (κ) were employed in statistical analysis of results.RESULTS:
In a first set of IHC reproducibility experiments, almost perfect agreement was recorded (κ=0.94) when two different operators scored CTC as overall positive or negative for AR. However, BCTI analysis identified a specific bias in scoring staining intensity, where one operator favoured moderate over strong assignments, whereas the reverse was the case with the second operator. After a period of additional training involving deployment of a panel of standardised images, a second set of validation experiments were conducted. These showed correction of the inter-operator bias by BCTI with κ for scoring intensity increasing from 0.59 to 0.81, indicative of almost perfect agreement.CONCLUSIONS:
By application of BCTI to the validation of IHC, operator bias and therefore poor reproducibility can be identified, characterised and corrected to achieve a level of error normally associated with a quantitative biomarker assay, such as an ELISA. The methodological approach described herein can be applied to any generic IHC technique.
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Base de dados:
MEDLINE
Assunto principal:
Piridazinas
/
Imunoquímica
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Receptores Androgênicos
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Células Neoplásicas Circulantes
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Antineoplásicos
Tipo de estudo:
Diagnostic_studies
/
Prognostic_studies
Limite:
Aged
/
Humans
/
Male
Idioma:
En
Ano de publicação:
2014
Tipo de documento:
Article