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1.
APMIS ; 131(5): 197-205, 2023 May.
Artigo em Inglês | MEDLINE | ID: mdl-36776120

RESUMO

Double-stranded RNA (dsRNA) is produced during most viral infections, and immunohistochemical detection of dsRNA has been proposed as a potential screening marker for viral replication. The anti-dsRNA monoclonal antibody clone 9D5 is more sensitive than the established clone J2 but has not been validated in formalin-fixed paraffin-embedded (FFPE) tissue. This study aimed to test and compare the performance of the anti-dsRNA monoclonal antibodies, 9D5 and J2, in FFPE tissue using an automated staining platform. Archived clinical tissue samples with viral infections (n = 34) and uninfected controls (n = 30) were examined. Immunohistochemical staining for dsRNA (9D5 and J2) and virus-specific epitopes was performed. 9D5 provided a similar staining pattern but a higher signal-to-noise ratio than J2. The following proportions of virus-infected tissue samples were dsRNA-positive: SARS-CoV-2 (5/5), HPV (6/6), MCV (5/5), CMV (5/6), HSV (4/6), and EBV (0/6). Also, 18 of 30 uninfected samples were dsRNA positive, and an association between fixation time and intensity was observed. However, signals in all samples were markedly reduced by pretreatment with dsRNA-specific RNAse-III, indicating a specific reaction. In conclusion, dsRNA can be demonstrated in most viral infections with immunohistochemistry in FFPE tissue but with low clinical specificity. The antibody clone 9D5 performs better than clone J2.


Assuntos
COVID-19 , Viroses , Humanos , RNA de Cadeia Dupla , Inclusão em Parafina , SARS-CoV-2 , Formaldeído
2.
Appl Immunohistochem Mol Morphol ; 31(7): 452-458, 2023 08 01.
Artigo em Inglês | MEDLINE | ID: mdl-36194495

RESUMO

Immunohistochemistry (IHC) has for decades been an integrated method within pathology applied to gain diagnostic, prognostic, and predictive information. However, the multimodality of the analytical phase of IHC is a challenge to ensure the reproducibility of IHC, which has been documented by external quality assessment (EQA) programs for many biomarkers. More than 600 laboratories participate in the Nordic immunohistochemical Quality Control EQA program for IHC. In the period, 2017-2021, 65 different biomarkers were assessed and a total of 31,967 results were evaluated. An overall pass rate of 79% was obtained being an improvement compared with 71% for the period, 2003-2015. The pass rates for established predictive biomarkers (estrogen receptor, progesterone receptor, and HER2) for breast carcinoma were most successful showing mean pass rates of 89% to 92%. Diagnostic IHC biomarkers as PAX8, SOX10, and different cytokeratins showed a wide spectrum of pass rates ranging from 37% to 95%, mean level of 75%, and attributed to central parameters as access to sensitive and specific antibodies but also related to purpose of the IHC test and validation performed accordingly to this. Seven new diagnostic biomarkers were introduced, and all showed inferior pass rates compared with the average level for diagnostic biomarkers emphasizing the challenge to optimize, validate, and implement new IHC biomarkers. Nordic immunohistochemical Quality Control operates by "Fit-For-Purpose" EQA principles and for programmed death-ligand 1, 2 segments are offered aligned to the "3-dimensional" approach-bridging diagnostic tests, drugs to be offered, and diseases addressed. Mean pass rates of 65% and 79% was obtained in the 2 segments for programmed death-ligand 1.


Assuntos
Neoplasias da Mama , Ensaio de Proficiência Laboratorial , Humanos , Feminino , Reprodutibilidade dos Testes , Imuno-Histoquímica , Ensaio de Proficiência Laboratorial/métodos , Controle de Qualidade , Receptores de Estrogênio , Neoplasias da Mama/diagnóstico , Biomarcadores Tumorais/análise , Receptor ErbB-2
3.
Appl Immunohistochem Mol Morphol ; 30(5): 326-332, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35510771

RESUMO

Immunohistochemistry for Uroplakin (UP) II and III is used to determine urothelial origin of carcinomas of unknown primary site and are especially valuable to differentiate urothelial carcinomas (UCs) from lung squamous cell carcinomas and prostate carcinomas. In the Nordic immunohistochemical Quality Control assessment scheme, only 45% of the participants obtained a sufficient staining result for UP. Primary antibodies (Abs) against UPII were most successful with a pass rate of 86%. No Abs against UPIII provided sufficient staining results. A comparative study was carried out on a larger cohort of tissue samples with optimized methods for the UPII mouse monoclonal antibody (mmAb) clone BC21, UPIII mmAb clone AU-1, and rabbit monoclonal antibody (rmAb) clone SP73 to evaluate the performance in a standardized way. Tissue microarrays containing 58 UCs, 111 non-UCs, and 20 normal tissues were included. The UP stains were evaluated by using H-score. Based on H-scores, samples were categorized as high-expressor (150 to 300), moderate-expressor (10 to 149), low-expressor (1 to 9), and negative (<1). The UPII mmAb clone BC21 obtained a significant higher analytical sensitivity of 69% for UCs compared with the UPIII Abs mmAb clone AU-1 and rmAb clone SP73 with 19% and 29%, respectively. No high-expressor UCs were seen for the UPIII Abs, whereas 13% of the positive UCs obtained an H-score >150 for the UPII Ab. The 2 UPIII Abs gave an analytical specificity of 100% compared with 97% for the UPII Ab being positive in 2 ovarian carcinomas and 1 cervical squamous cell carcinoma.


Assuntos
Carcinoma de Células Escamosas , Carcinoma de Células de Transição , Neoplasias da Bexiga Urinária , Animais , Anticorpos Monoclonais , Carcinoma de Células Escamosas/diagnóstico , Carcinoma de Células Escamosas/patologia , Carcinoma de Células de Transição/diagnóstico , Carcinoma de Células de Transição/patologia , Feminino , Humanos , Masculino , Camundongos , Neoplasias da Bexiga Urinária/diagnóstico , Neoplasias da Bexiga Urinária/patologia , Uroplaquina II , Urotélio/patologia
4.
Front Neurosci ; 16: 836259, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35360166

RESUMO

Modern diffusion and functional magnetic resonance imaging (dMRI/fMRI) provide non-invasive high-resolution images from which multi-layered networks of whole-brain structural and functional connectivity can be derived. Unfortunately, the lack of observed correspondence between the connectivity profiles of the two modalities challenges the understanding of the relationship between the functional and structural connectome. Rather than focusing on correspondence at the level of connections we presently investigate correspondence in terms of modular organization according to shared canonical processing units. We use a stochastic block-model (SBM) as a data-driven approach for clustering high-resolution multi-layer whole-brain connectivity networks and use prediction to quantify the extent to which a given clustering accounts for the connectome within a modality. The employed SBM assumes a single underlying parcellation exists across modalities whilst permitting each modality to possess an independent connectivity structure between parcels thereby imposing concurrent functional and structural units but different structural and functional connectivity profiles. We contrast the joint processing units to their modality specific counterparts and find that even though data-driven structural and functional parcellations exhibit substantial differences, attributed to modality specific biases, the joint model is able to achieve a consensus representation that well accounts for both the functional and structural connectome providing improved representations of functional connectivity compared to using functional data alone. This implies that a representation persists in the consensus model that is shared by the individual modalities. We find additional support for this viewpoint when the anatomical correspondence between modalities is removed from the joint modeling. The resultant drop in predictive performance is in general substantial, confirming that the anatomical correspondence of processing units is indeed present between the two modalities. Our findings illustrate how multi-modal integration admits consensus representations well-characterizing each individual modality despite their biases and points to the importance of multi-layered connectomes as providing supplementary information regarding the brain's canonical processing units.

5.
Appl Immunohistochem Mol Morphol ; 29(9): 643-647, 2021 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-34545849

RESUMO

This paper is number 8 in a series developed through a partnership between ISIMM and NordiQC with the purpose of reporting research assessing the performance characteristics of immunoassays in an external proficiency testing program.


Assuntos
Ensaio de Proficiência Laboratorial , Controle de Qualidade , Sinaptofisina/metabolismo , Humanos , Imunoensaio
6.
Cancers (Basel) ; 13(5)2021 Mar 09.
Artigo em Inglês | MEDLINE | ID: mdl-33803148

RESUMO

We compared estrogen receptor (ER), progesterone receptor (PR), human epidermal growth-factor receptor 2 (HER2), Ki67, and grade scores among the pathology departments in Sweden. We investigated how ER and HER2 positivity rates affect the distribution of endocrine and HER2-targeted treatments among oncology departments. All breast cancer patients diagnosed between 2013 and 2018 in Sweden were identified in the National Quality Register for Breast Cancer. Cases with data on ER, PR, HER2, Ki67, grade, and treatment were selected (43,261 cases from 29 departments following the guidelines for biomarker testing). The ER positivity rates ranged from 84.2% to 97.6% with 6/29 labs out of the overall confidence intervals (CIs), while PR rates varied between 64.8% and 86.6% with 7/29 labs out of the CIs. HER2 positivity rates ranged from 9.4% to 16.3%, with 3/29 labs out of the overall CIs. Median Ki67 varied between 15% and 30%, where 19/29 labs showed significant intra-laboratory variability. The proportion of grade-II cases varied between 42.9% and 57.1%, and 13/29 labs were outside of the CI. Adjusting for patient characteristics, the proportion of endocrine and anti-HER2 treatments followed the rate of ER and HER2 positivity, illustrating the clinical effect of inter- and intra-laboratory variability. There was limited variability among departments in ER, PR, and HER2 testing. However, even a few outlier pathology labs affected endocrine and HER2-targeted treatment rates in a clinically relevant proportion, suggesting the need for improvement. High variability was found in grading and Ki67 assessment, illustrating the need for the adoption of new technologies in practice.

8.
Appl Immunohistochem Mol Morphol ; 29(2): 99-104, 2021 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-32168036

RESUMO

Immunohistochemical staining for Ki-67 is used to calculate a Ki-67 proliferation index (PI) that carries prognostic and predictive information in various cancers including breast carcinomas. Studies have documented challenges for observers to reproducibly estimate the Ki-67 PI. At present, no international consensus exists concerning scoring method (eg, hotspots vs. overall average, digital vs. manual counting) or even the definition of a Ki-67-positive cell. To clarify the approach to Ki-67 scoring and evaluation of the interobserver agreement among participants in the Nordic Immunohistochemical Quality Control (NordiQC) Breast Cancer Module, a study was set up on the basis of an online web module containing 15 digitized tissue microarray cores of breast carcinomas stained for Ki-67 in the NordiQC reference laboratory. All participants were invited to attend the study. In addition to Ki-67 scoring, they were asked to disclose their preferred method for Ki-67 estimation and their job title. For comparison, slides were analyzed using a Digital Image Analysis algorithm based on Virtual Double Staining. In total, 199 participants enrolled for the study. Overall, there was a good correlation in Ki-67 PIs among the participants, although results for some cores varied significantly. However, when applying a cutoff of 20%, a relatively low κ value of 0.52 was observed. Participants scoring in hotspots reported higher Ki-67 PIs than participants estimating an overall average, and, not surprisingly, participants who considered weak Ki-67 nuclear staining positive obtained higher Ki-67 PIs than those who did not. Ki-67 PI was not correlated to job title. The Virtual Double Staining algorithm obtained Ki-67 values close to the mean value of the human observers. Our study underlines the need for international standardization and guidelines in estimation of Ki-67 PI. Digital Image Analysis may be a useful tool in this process.


Assuntos
Algoritmos , Neoplasias da Mama , Processamento de Imagem Assistida por Computador , Antígeno Ki-67/metabolismo , Índice Mitótico , Coloração e Rotulagem , Adulto , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Proliferação de Células , Feminino , Humanos , Pessoa de Meia-Idade
9.
Appl Immunohistochem Mol Morphol ; 28(7): 566-570, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32243261

RESUMO

This paper is number 7 in a series developed through a partnership between ISIMM and NordiQC with the purpose of reporting research assessing the performance characteristics of immunoassays in an external proficiency testing program.


Assuntos
Anticorpos Monoclonais/imunologia , Células Epiteliais/metabolismo , Imuno-Histoquímica/métodos , Queratina-5/metabolismo , Ensaio de Proficiência Laboratorial/métodos , Animais , Humanos , Cininogênio de Alto Peso Molecular , Controle de Qualidade
10.
Mod Pathol ; 33(1): 4-17, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31383961

RESUMO

Different clones, protocol conditions, instruments, and scoring/readout methods may pose challenges in introducing different PD-L1 assays for immunotherapy. The diagnostic accuracy of using different PD-L1 assays interchangeably for various purposes is unknown. The primary objective of this meta-analysis was to address PD-L1 assay interchangeability based on assay diagnostic accuracy for established clinical uses/purposes. A systematic search of the MEDLINE database using PubMed platform was conducted using "PD-L1" as a search term for 01/01/2015 to 31/08/2018, with limitations "English" and "human". 2,515 abstracts were reviewed to select for original contributions only. 57 studies on comparison of two or more PD-L1 assays were fully reviewed. 22 publications were selected for meta-analysis. Additional data were requested from authors of 20/22 studies in order to enable the meta-analysis. Modified GRADE and QUADAS-2 criteria were used for grading published evidence and designing data abstraction templates for extraction by reviewers. PRISMA was used to guide reporting of systematic review and meta-analysis and STARD 2015 for reporting diagnostic accuracy study. CLSI EP12-A2 was used to guide test comparisons. Data were pooled using random-effects model. The main outcome measure was diagnostic accuracy of various PD-L1 assays. The 22 included studies provided 376 2×2 contingency tables for analyses. Results of our study suggest that, when the testing laboratory is not able to use an Food and Drug Administration-approved companion diagnostic(s) for PD-L1 assessment for its specific clinical purpose(s), it is better to develop a properly validated laboratory developed test for the same purpose(s) as the original PD-L1 Food and Drug Administration-approved immunohistochemistry companion diagnostic, than to replace the original PD-L1 Food and Drug Administration-approved immunohistochemistry companion diagnostic with a another PD-L1 Food and Drug Administration-approved companion diagnostic that was developed for a different purpose.


Assuntos
Antígeno B7-H1/análise , Imuno-Histoquímica/métodos , Humanos , Imuno-Histoquímica/normas
11.
Cytometry A ; 95(10): 1066-1074, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31490627

RESUMO

Bone marrow cellularity is an important measure in diagnostic hematopathology. Currently, the gold standard for bone marrow cellularity estimation is manual inspection of hematoxylin and eosin stained whole slide images (H&E WSI) by hematopathologists. However, these assessments are subjective and subject to interobserver and intraobserver variability. This may be reduced by using a computer-assisted estimate of bone marrow cellularity. The aim of this study was to develop a fully automated algorithm to estimate bone marrow cellularity in H&E WSI stains using bone marrow segmentation. Data consisted of eight bone marrow H&E WSIs extracted from eight subjects. An algorithm was developed to estimate the bone marrow cellularity consisting of biopsy segmentation, tissue classification, and bone marrow segmentation. Segmentations of the red and yellow bone marrow (YBM) were used to estimate the bone marrow cellularity within the WSI H&E stains. The DICE coefficient between automatic tissue segmentations and ground truth segmentations conducted by an experienced hematopathologist were used for validation. Furthermore, the agreement between the automatic and two manual cellularity estimates was assessed using Bland-Altman plots and intraclass correlation coefficients (ICC). The validation of the bone marrow segmentation demonstrated an average DICE of 0.901 and 0.920 for the red and YBM, respectively. A mean cellularity estimate difference of -0.552 and - 7.816 was obtained between the automatic cellularity estimates and two manual cellularity estimates, respectively. An ICC of 0.980 (95%CI: 0.925-0.995, P-value: 5.51 × 10-7 ) was obtained between the automatic and manual cellularity estimates based on manual annotations. The study demonstrated that it was possible to obtain bone marrow cellularity estimates with a good agreement with bone marrow cellularity estimates obtained from an experienced hematopathologist. © 2019 International Society for Advancement of Cytometry.


Assuntos
Células da Medula Óssea/citologia , Processamento de Imagem Assistida por Computador , Coloração e Rotulagem , Algoritmos , Automação , Humanos
12.
Appl Immunohistochem Mol Morphol ; 27(10): 732-739, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31460869

RESUMO

Ki67 is a nuclear protein expressed during the active phases of the cell cycle, which makes it a biomarker of cell proliferation. In clinical pathology settings, immunohistochemical (IHC) detection of Ki67 is used to calculate Ki67 proliferation indices (PIs), which have prognostic information and are used to subdivide breast carcinomas and neuroendocrine neoplasias. Calculation of Ki67 PIs is notoriously hard and prone to intraobserver and interobserver variance. In addition, IHC protocol settings [such as primary antibody (Ab) clone, clone format, and stainer platform] can affect the result of the IHC assays and in turn the Ki67 PI. Digital image analysis has been suggested as a useful tool to standardize Ki67 counting. Recently, virtual double staining, a computer algorithm segmenting Ki67 and Ki67 tumor cells using digitally fused parallel cytokeratin and Ki67-stained slides, has been introduced. In this study, we compare Ki67 PIs obtained by virtual double staining in 41 breast carcinomas stained using the most commonly used commercially available primary Ab clones and formats on the main stainer platforms. IHC protocols for the concentrated (conc) Ab and platform combinations were optimized for the highest analytical sensitivity and optimal signal-to-noise ratio, whereas ready-to-use (RTU) formats were used, as recommended by the vendor. Significant differences in the mean Ki67 PIs (relativized to the mean core Ki67) were observed not only between the different Ab clones but also the different formats and stainer platforms; Ki67 PIs with SP6 conc stained on the Ventana BenchMark ULTRA platform were on average 11.9 percentage points (pp) higher than the mean core average, whereas with Ab 30.9 RTU on the Ventana platform, they were 10.4 pp higher. Mib1 RTU (Dako Autostainer Link 48) and MM1 RTU (Leica Bond) provided 8.6 and 12.5 pp lower Ki67 PIs, respectively. Mib1 conc and SP6 conc on the Dako Autostainer and Leica Bond provided similar results-close to the overall average. Significant variations in the proportion of tumors with Ki67 high-level expression (Ki67 PI ≥20%) were observed among Ab, format, and stainer platform combinations. The results underline the challenges in the comparison of Ki67 PIs across Abs, formats, and platforms. Researchers and clinicians need to account for these differences when reporting Ki67 PIs. To advance the usefulness of Ki67 PIs in the research and clinical setting, standardization of Ki67 IHC assays is needed.


Assuntos
Biomarcadores/metabolismo , Neoplasias da Mama/diagnóstico , Carcinoma Ductal/diagnóstico , Diagnóstico por Imagem/métodos , Antígeno Ki-67/metabolismo , Tumores Neuroendócrinos/diagnóstico , Anticorpos Monoclonais/metabolismo , Proliferação de Células , Células Clonais , Diagnóstico Diferencial , Diagnóstico por Imagem/normas , Feminino , Humanos , Imuno-Histoquímica , Pessoa de Meia-Idade , Índice Mitótico , Prognóstico , Sensibilidade e Especificidade , Coloração e Rotulagem
13.
Case Rep Pathol ; 2019: 2734815, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31198613

RESUMO

Mediastinal pure choriocarcinomas are exceedingly rare representations of germ cell tumours and are associated with a poor prognosis. To date, fewer than 20 cases have been reported. This current report describes an elderly patient who developed a large rapidly growing mediastinal tumour. Unfortunately, the patient expired before a definitive diagnosis could be reached. An autopsy revealed that the histomorphological features of the tumour showed two distinct tumour cell populations (syncytio- and cytotrophoblasts), and the diagnosis of choriocarcinoma was made. Immunohistochemical analysis showed a characteristic staining pattern in agreement with published studies. Here, we report a case of primary mediastinal choriocarcinoma in an elderly male with concurrent metastasizing prostate adenocarcinoma treated with long-term goserelin deposits, which, as we speculate, could have induced the choriocarcinoma.

14.
Hear Res ; 378: 176-184, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30826128

RESUMO

BACKGROUND: Middle ear physiology includes both sound pressure transmission and homeostasis of its static air pressure. Pressure gradients are continuously created by gas exchange over the middle ear mucosa as well as by ambient pressure variations. Gas exchange models require actual values for regional mucosa thickness, blood vessel density, and diffusion distance. Such quantitative data have been scarce and limited to few histological samples from the tympanic cavity (TC) and the antrum. However, a detailed regional description of the morphological differences of the TC and mastoid air cell system (MACS) mucosa has not been available. The aim of the present study was to provide such parameters. METHODS: The study included sets of three histological H&E-slides from 15 archived healthy temporal bones. We performed a comparison of the mucosa morphology among the following regions: (1) anterior TC; (2) inferior TC; (3) posterior TC; (4) superior TC; (5) MACS antrum; (6) superior MACS; (7) central MACS; (8) inferior MACS. RESULTS: Regions (1)-(3), situated below the inter-attico-tympanic diaphragm, had the largest proportion of high respiratory epithelium, cilia and loose lamina propria within the mucosa, as well as the thickest mucosa and the largest diffusion distance. Regions (6)-(8), situated above the diaphragm, had the thinnest mucosa, the shortest distance to the blood vessels, together with the largest proportion of flat epithelium and very few cilia. Regions (4)-(5), still supradiaphragmatic, had intermediary values for these parameters, but generally closer to regions (6)-(8). The blood vessel density and the proportion of active mucosa were not significantly different among the regions. CONCLUSION: Mucosa of regions (1), (2) and (3) represented a predominantly clearance-specific morphology, whereas in regions (4)-(8) it seemed adapted to gas exchange. However, the lack of statistically significant differences in blood vessel density and proportion of active mucosa indicated that all regions could be involved in gas exchange with the highest adaptation in the superior MACS. This pattern divides the middle ear functionally along the inter-attico-tympanic diaphragm rather than the anatomical division into TC and MACS.


Assuntos
Orelha Média/fisiologia , Audição , Mucosa Respiratória/fisiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Pressão do Ar , Cadáver , Difusão , Orelha Média/anatomia & histologia , Feminino , Homeostase , Humanos , Masculino , Pessoa de Meia-Idade , Movimento (Física) , Mucosa Respiratória/anatomia & histologia , Som , Adulto Jovem
15.
Oncotarget ; 10(7): 717-731, 2019 Jan 22.
Artigo em Inglês | MEDLINE | ID: mdl-30774774

RESUMO

Survival of diffuse large B-cell lymphoma (DLBCL) patients has improved by inclusion of rituximab. Refractory/recurrent disease caused by treatment resistance is, however, a major problem. Determinants of rituximab sensitivity are not fully understood, but effect of rituximab are enhanced by antagonizing cell surface receptor CXCR4. In a two-step strategy, we tested the hypothesis that prognostic value of CXCR4 in DLBCL relates to rituximab treatment, due to a hampering effect of CXCR4 on the response of DLBCL cells to rituximab. First, by investigating the prognostic impact of CXCR4 mRNA expression separately for CHOP (n=181) and R-CHOP (n=233) cohorts and, second, by assessing the interaction between CXCR4 and rituximab in DLBCL cell lines. High CXCR4 expression level was significantly associated with poor outcome only for R-CHOP-treated patients, independent of IPI score, CD20 expression, ABC/GCB and B-cell-associated gene signature (BAGS) classifications. s. For responsive cell lines, inverse correlation was observed between rituximab sensitivity and CXCR4 surface expression, rituximab induced upregulation of surface-expressed CXCR4, and growth-inhibitory effect of rituximab increased by plerixafor, supporting negative impact of CXCR4 on rituximab function. In conclusion, CXCR4 is a promising independent prognostic marker for R-CHOP-treated DLBCL patients, possibly due to inverse correlation between CXCR4 expression and rituximab sensitivity.

16.
Appl Immunohistochem Mol Morphol ; 27(4): 258-262, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30653032

RESUMO

This paper is number 6 in a series developed through a partnership between ISIMM and Nordic IHC Quality Control with the purpose of reporting research assessing the performance characteristics of immunoassays in an external proficiency testing program.


Assuntos
Anticorpos Monoclonais/química , Cromogranina A/metabolismo , Técnicas de Laboratório Clínico/normas , Ensaio de Proficiência Laboratorial , Humanos , Imunoensaio
17.
Appl Immunohistochem Mol Morphol ; 27(2): 87-91, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30499819

RESUMO

This paper is the number 5 in a series developed through a partnership between ISIMM and NordiQC for the purpose of reporting research assessing the performance characteristics of immunoassays in an external proficiency-testing program.


Assuntos
Neoplasias Gastrointestinais/metabolismo , Células-Tronco Hematopoéticas/metabolismo , Imunoensaio/métodos , Ensaio de Proficiência Laboratorial/métodos , Proteínas Proto-Oncogênicas c-kit/metabolismo , Dinamarca , Testes Diagnósticos de Rotina , Neoplasias Gastrointestinais/diagnóstico , Humanos , Garantia da Qualidade dos Cuidados de Saúde , Controle de Qualidade
18.
Cytometry A ; 95(4): 381-388, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30556331

RESUMO

Breast cancer is the most frequent cancer among women worldwide. Ki67 can be used as an immunohistochemical pseudo marker for cell proliferation to determine how aggressive the cancer is and thereby the treatment of the patient. No standard Ki67 staining protocol exists, resulting in inter-laboratory stain variability. Therefore, it is important to determine the quality control of a staining protocol to ensure correct diagnosis and treatment of patients. Currently, quality control is performed by the organization NordiQC that use an expert panel-based qualitative assessment system. However, no objective method exists to determine the quality of a staining protocol. In this study, we propose an algorithm, to objectively assess staining quality from segmented cell nuclei structures extracted from cell lines. The cell nuclei were classified into either Ki67 positive or negative to determine the Ki67 proliferation index within the cell lines. A Ki67 stain quality model based on ordinal logistic regression was developed to determine the quality of a staining protocol from features extracted from the segmented cell nuclei in the cell lines. The algorithm was able to segment and classify Ki67 positive cell nuclei with a sensitivity and positive predictive value (PPV) of 0.90 and 0.94 and Ki67 negative cell nuclei with a sensitivity and PPV of 0.78 and 0.78. The mean difference between a manual and automatic Ki67 proliferation index was -0.003 with a standard deviation of 0.056. The ordinal logistic regression model found that the stain intensity for both the Ki67 positive and Ki67 negative cell nuclei were statistically significant as parameters determining the stain quality from the cell line cores. The framework shows great promise for using cell nuclei information from cell lines to predict the staining quality of staining protocols. © 2018 International Society for Advancement of Cytometry.


Assuntos
Algoritmos , Proliferação de Células , Processamento de Imagem Assistida por Computador , Antígeno Ki-67/metabolismo , Controle de Qualidade , Coloração e Rotulagem/normas , Biomarcadores Tumorais/análise , Biomarcadores Tumorais/metabolismo , Neoplasias da Mama/diagnóstico , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Núcleo Celular/metabolismo , Feminino , Humanos , Processamento de Imagem Assistida por Computador/métodos , Processamento de Imagem Assistida por Computador/normas , Índice Mitótico , Prognóstico , Coloração e Rotulagem/métodos
19.
Appl Immunohistochem Mol Morphol ; 26(7): 431-434, 2018 08.
Artigo em Inglês | MEDLINE | ID: mdl-30024425

RESUMO

This paper is number 4 in a series developed through a partnership between ISIMM and Nordic immunohistochemical Quality Control for the purpose of reporting research assessing the performance characteristics of immunoassays in an external proficiency testing program.


Assuntos
Proteínas de Ligação a DNA/metabolismo , Controle de Qualidade , Proteínas de Ligação a DNA/imunologia , Humanos , Imunoensaio/métodos , Imunoensaio/normas
20.
Appl Immunohistochem Mol Morphol ; 26(4): 221-224, 2018 04.
Artigo em Inglês | MEDLINE | ID: mdl-29621095

RESUMO

This paper is number 3 in a series developed through a partnership between ISIMM and NordiQC for the purpose of reporting research assessing the performance characteristics of immunoassays in an external proficiency testing program.


Assuntos
Imunoensaio/métodos , Imuno-Histoquímica/métodos , Fator de Transcrição PAX8/metabolismo , Automação Laboratorial , Técnicas de Laboratório Clínico , Reações Cruzadas , Humanos , Ensaio de Proficiência Laboratorial , Variações Dependentes do Observador , Fator de Transcrição PAX8/imunologia , Controle de Qualidade
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