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2. Comparison of different methodologies and cryostat versus paraffin sections for chromogenic immunohistochemistryVashendriya V. V. Hira, Annique Loncq de Jong, Klea Ferro, Mohammed Khurshed, Remco J. Molenaar, Cornelis J. F. van Noorden, 2019, original scientific article Abstract: Immunohistochemistry (IHC) specifically localizes proteins in cells and tissues, but methodologies vary widely. Therefore, we performed a methodological IHC optimization and validation study. First, we compared advantages and disadvantages of cryostat sections versus paraffin sections. Second, we compared and optimized antigen retrieval in paraffin sections using citrate buffer and Tris/EDTA buffer. Third, aminoethyl carbazole (AEC) and 3,3'-diaminobenzidine (DAB) were tested as horseradish peroxidase (HRP) substrates to obtain a water-insoluble coloured end product to visualize antigens. Fourth, secondary antibodies conjugated with either mono-HRP or poly-HRP were compared. The study was performed using serial sections of human tonsil. IHC was performed with primary antibodies against endothelial cell marker CD31, smooth muscle actin (SMA), chemokine stromal-derived factor-1α (SDF-1α) and its receptor C-X-C receptor type 4 (CXCR4), macrophage marker CD68 and proliferation marker Ki67. DAB rather than AEC, and cryostat sections rather than paraffin sections gave optimum staining at highest primary antibody dilutions, whereas tissue morphology in paraffin sections was superior. Loss of antigenicity in paraffin sections by formaldehyde fixation, heat and/or masking of epitopes was counteracted by antigen retrieval but not for all antigens. Two out of six antigens (CD31 and CD68) could not be retrieved irrespective time and type of retrieval. Tris-EDTA was superior to citrate buffer for antigen retrieval. The use of mono-HRP or poly-HRP depended on the affinity of the primary antibody for its antigen. We conclude that IHC methodology optimization and validation are crucial steps for each antibody and each research question.
Keywords: immunohistochemistry, chromogens, aminoethyl carbazole, AEC, 3, 3'-diaminobenzidine, DAB, antigen retrieval, tonsil
Published in DiRROS: 24.07.2024; Views: 102; Downloads: 94
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3. Development and characterization of a novel mAb against bilitranslocase - a new biomarker of renal carcinomaSendi Montanič, Michela Terdoslavich, Uroš Rajčević, Luigina De Leo, Serena Bonin, Vladka Čurin-Šerbec, Sabina Passamonti, 2013, original scientific article Keywords: bilitranslocase, monoclonal antibody, peptide antigen
Published in DiRROS: 22.03.2024; Views: 250; Downloads: 73
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5. Development of the recycling procedure for rapid antigen testsRebeka Rudolf, Darja Feizpour, Žiga Jelen, Peter Majerič, Tilen Švarc, Matej Zadravec, Timi Gomboc, Aleksandra Kocijan, 2024, original scientific article Keywords: rapid antigen tests, recycling, characterization, nanogold, plastic
Published in DiRROS: 28.02.2024; Views: 391; Downloads: 152
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6. Automated analysis of proliferating cells spatial organisation predicts prognosis in lung neuroendocrine neoplasmsMatteo Bulloni, Giada Sandrini, Irene Stacchiotti, M. Barberis, Fiorella Calabrese, Lina Carvalho, Gabriella Fontanini, Greta Alì, Francesco Fortarezza, Paul Hofman, Izidor Kern, 2021, original scientific article Keywords: neuroendocrine tumors, lung neoplasms, carcinoma, prognosis, pathology, histology, Ki-67 antigen, machine learning, image enhancement, lung cancer, neuroendocrine neoplasms
Published in DiRROS: 09.11.2021; Views: 990; Downloads: 425
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7. Usefulness of rapid antigen testing for SARS-CoV-2 screening of healthcare workers : ǂa ǂpilot studyAnja Šterbenc, Viktorija Tomič, Urška Bidovec, Katja Vrankar, Aleš Rozman, Mihaela Zidarn, 2021, other scientific articles Abstract: Background. Identification of infected healthcare workers (HCWs) is an important step in severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) transmission control. Rapid antigen tests (RATs) are considered an important addition to molecular tests in diagnosing coronavirus disease 2019 (COVID-19), mainly because of their fast turnaround time, easier analytical procedure and lower price. However, real-life studies on the usefulness of such testing for screening of HCWs are limited. Methods. Physicians, nurses and hospital attendants currently working at the University Clinic of Respiratory and Allergic Diseases Golnik were invited to participate in the pilot study. Nasopharyngeal swabs were obtained three times per week for two consecutive weeks and tested with a point-of-care RAT and reverse transcription polymerase chain reaction (RT-PCR). Serum samples were obtained at the beginning of the study and 2 weeks after the last swab was collected to evaluate the serological status. Results. A total of 191 nasopharyngeal swabs from 36 HCWs were obtained. None of the samples tested was positive for the presence of SARS-CoV-2 antigen, whereas two HCWs tested positive on RT-PCR. Of these, one HCW had a newly identified SARS-CoV-2 infection, whereas RT-PCR probably detected a previous but recent infection in the other HCW. Conclusio.n Based on the results of this pilot study, it is unlikely that RAT will reliably detect novel SARS-CoV-2 infections among asymptomatic HCWs despite serial sampling. Although RT-PCR-based screening of HCWs may not be feasible due to high sample volume, molecular methods may identify SARS-CoV-2-infected HCWs already during the presymptomatic stage.
Keywords: SARS-CoV-2, health personnel, COVID-19 serological testing, real-time polymerase chain reaction, rapid antigen test, screening
Published in DiRROS: 28.05.2021; Views: 1237; Downloads: 381
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