Formalin, an aqueous solution of formaldehyde, has been the gold standard for fixation of histological samples for over a century. Despite its considerable advantages, growing evidence points to objective toxicity, particularly highlighting its carcinogenicity and mutagenic effects. In 2016, the European Union proposed a ban, but a temporary permission was granted in consideration of its fundamental role in the medical-diagnostic field. In the present study, we tested an innovative fixative, glyoxal acid-free (GAF) (a glyoxal solution deprived of acids), which allows optimal tissue fixation at structural and molecular level combined with the absence of toxicity and carcinogenic activity. An open-label, non-inferiority, multicentric trial was performed comparing fixation of histological specimens with GAF fixative vs standard phosphate-buffered formalin (PBF), evaluating the morphological preservation and the diagnostic value with four binary score questions answered by both the central pathology reviewer and local center reviewers. The mean of total score in the GAF vs PBF fixative groups was 3.7 ± 0.5 vs 3.9 ± 0.3 for the central reviewer and 3.8 ± 0.5 vs 4.0 ± 0.1 for the local pathologist reviewers, respectively. In terms of median value, similar results were observed between the two fixative groups, with a median value of 4.0. Data collected indicate the non-inferiority of GAF as compared to PBF for all organs tested. The present clinical performance study, performed following the international standard for performance evaluation of in vitro diagnostic medical devices, highlights the capability of GAF to ensure both structural preservation and diagnostic value of the preparations.
- Klíčová slova
- test MammaPrint,
- MeSH
- diagnostické techniky molekulární metody přístrojové vybavení MeSH
- fixace tkání MeSH
- genetické testování * metody přístrojové vybavení MeSH
- klinické laboratoře organizace a řízení zákonodárství a právo MeSH
- klinické zkoušky, fáze III jako téma MeSH
- lidé MeSH
- metody pro přípravu analytických vzorků MeSH
- nádory prsu * diagnóza genetika MeSH
- staging nádorů MeSH
- stanovení celkové genové exprese metody MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- novinové články MeSH
The dysplasia grading of Barrett's esophagus (BE), based on the histomorphological assessment of formalin-fixed, paraffin-embedded (FFPE) tissue, suffers from high interobserver variability leading to an unsatisfactory prediction of cancer risk. Thus, pre-analytic preservation of biological molecules, which could improve risk prediction in BE enabling molecular and genetic analysis, is needed. We aimed to evaluate such a molecular pre-analytic fixation tool, PAXgene-fixed paraffin-embedded (PFPE) biopsies, and their suitability for histomorphological BE diagnostics in comparison to FFPE. In a ring trial, 9 GI pathologists evaluated 116 digital BE slides of non-dysplastic BE (NDBE), low-grade dysplasia (LGD), high-grade dysplasia (HGD), and esophageal adenocarcinomas (EAC) using virtual microscopy. Overall quality, cytological and histomorphological parameters, dysplasia criteria, and diagnosis were analyzed. PFPE showed better preservation of nuclear details as chromatin and nucleoli, whereas overall quality and histomorphologic parameters as visibility of basal lamina, goblet cells, and presence of artifacts were scored as equal to FFPE. The interobserver reproducibility with regard to the diagnosis was best for NDBE and EAC (κF = 0.72-0.75) and poor for LGD and HGD (κF = 0.13-0.3) in both. In conclusion, our data suggest that PFPE allows equally confident histomorphological diagnosis of BE and EAC, introducing a novel tool for molecular analysis and parallel histomorphological evaluation.
- MeSH
- adenokarcinom * diagnóza genetika patologie MeSH
- Barrettův syndrom * diagnóza patologie MeSH
- fixace tkání MeSH
- hyperplazie MeSH
- lidé MeSH
- nádory jícnu * diagnóza patologie MeSH
- prekancerózy * patologie MeSH
- progrese nemoci MeSH
- reprodukovatelnost výsledků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- klinická studie MeSH
- Klíčová slova
- ozařovací fixační podprsenka,
- MeSH
- fixace tkání * metody MeSH
- kožní manifestace MeSH
- lidé MeSH
- nádory prsu radioterapie MeSH
- pronační poloha fyziologie MeSH
- prsy * anatomie a histologie účinky záření MeSH
- radioterapie přístrojové vybavení škodlivé účinky MeSH
- zdravotnické prostředky MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- MeSH
- fixace fraktury metody MeSH
- fixace tkání metody MeSH
- fraktury kostí diagnóza rehabilitace MeSH
- kineziologie aplikovaná * metody MeSH
- kosterní svaly fyziologie inervace růst a vývoj zranění MeSH
- lidé MeSH
- poranění ramene * chirurgie etiologie rehabilitace MeSH
- poranění šlachy diagnóza rehabilitace MeSH
- ramenní kloub anatomie a histologie patofyziologie růst a vývoj MeSH
- rozsah kloubních pohybů fyziologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- přehledy MeSH
Během září a října roku 2021 proběhl na popud Centra pro trombotické mikroangiopatie Ostrava (C4TMO) a České skupiny pro trombotické mikroangiopatie (CS4TMA) formou on-line dotazování připraveného společností IQVIA podle podkladů CS4TMA mezi klinickými hematology průzkum s cílem ověřit povědomí klinických hematologů o pravidlech a dostupnosti péče o pacienty s trombotickými mikroangiopatiemi. Odborným základem pro vytvoření sady otázek byly formulace pravidel správné klinické praxe péče o pacienty s trombotickou trombocytopenickou purpurou. Celkem bylo provedeno 49 rozhovorů. Výsledky průzkumu svědčí o velmi dobré klinické praxi péče o pacienty s akutní trombotickou mikroangiopatií na pracovištích respondentů. Spektrum pracovišť, které se zúčastnily průzkumu však neodpovídají skutečnému spektru hematologických pracovišť v České republice. Chybí data především z malých ambulancí v rámci nemocnic II. typu nebo mimo nemocnice.
From September to October 2021, at the request of the Centre for Thrombotic Microangiopathies Ostrava (C4TMO) and the Czech Group for Thrombotic Microangiopathies (CS4TMA), an online survey prepared by IQVIA based on CS4TMA data was conducted among clinical haematologists to verify the awareness of clinical haematologists about the rules and availability of care for patients with thrombotic microangiopathies. The professional basis for the development of the questionnaire was the formulation of good clinical practice guidelines for the care of patients with thrombotic thrombocytopenic purpura. A total of 49 interviews were conducted. The results of the survey indicate very good clinical practice for the care of patients with acute thrombotic microangiopathy at the respondents‘ institutions. However, the spectrum of departments that participated in the survey does not correspond to the actual spectrum of haematology departments in the Czech Republic. In particular, data from small outpatient clinics within type II hospitals or outside hospitals are missing.
- MeSH
- fixace tkání MeSH
- lidé MeSH
- protein ADAMTS13 krev MeSH
- průzkumy a dotazníky MeSH
- trombotické mikroangiopatie * diagnóza terapie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
Biomarker testing is crucial for treatment selection in advanced non-small cell lung cancer (NSCLC). However, the quantity of available tissue often presents a key constraint for patients with advanced disease, where minimally invasive tissue biopsy typically returns small samples. In Part 1 of this two-part series, we summarise evidence-based recommendations relating to small sample processing for patients with NSCLC. Generally, tissue biopsy techniques that deliver the greatest quantity and quality of tissue with the least risk to the patient should be selected. Rapid on-site evaluation can help to ensure sufficient sample quality and quantity. Sample processing should be managed according to biomarker testing requirements, because tissue fixation methodology influences downstream nucleic acid, protein and morphological analyses. Accordingly, 10% neutral buffered formalin is recommended as an appropriate fixative, and the duration of fixation is recommended not to exceed 24-48 h. Tissue sparing techniques, including the 'one biopsy per block' approach and small sample cutting protocols, can help preserve tissue. Cytological material (formalin-fixed paraffin-embedded [FFPE] cytology blocks and non-FFPE samples such as smears and touch preparations) can be an excellent source of nucleic acid, providing either primary or supplementary patient material to complete morphological and molecular diagnoses. Considerations on biomarker testing, reporting and quality assessment are discussed in Part 2.
- MeSH
- biologické markery MeSH
- fixace tkání metody MeSH
- fixativa MeSH
- formaldehyd MeSH
- lidé MeSH
- nádory plic * diagnóza patologie MeSH
- nemalobuněčný karcinom plic * diagnóza patologie MeSH
- nukleové kyseliny * MeSH
- zalévání tkání do parafínu MeSH
- znalecký posudek MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
BACKGROUND: Clinical laboratories routinely use formalin-fixed paraffin-embedded (FFPE) tissue or cell block cytology samples in oncology panel sequencing to identify mutations that can predict patient response to targeted therapy. To understand the technical error due to FFPE processing, a robustly characterized diploid cell line was used to create FFPE samples with four different pre-tissue processing formalin fixation times. A total of 96 FFPE sections were then distributed to different laboratories for targeted sequencing analysis by four oncopanels, and variants resulting from technical error were identified. RESULTS: Tissue sections that fail more frequently show low cellularity, lower than recommended library preparation DNA input, or target sequencing depth. Importantly, sections from block surfaces are more likely to show FFPE-specific errors, akin to "edge effects" seen in histology, while the inner samples display no quality degradation related to fixation time. CONCLUSIONS: To assure reliable results, we recommend avoiding the block surface portion and restricting mutation detection to genomic regions of high confidence.
Epitachophoresis is a novel next generation extraction system capable of isolating DNA and RNA simultaneously from clinically relevant samples. Here we build on the versatility of Epitachophoresis by extracting diverse nucleic acids ranging in lengths (20 nt-290 Kbp). The quality of extracted miRNA, mRNA and gDNA was assessed by downstream Next-Generation Sequencing.
- MeSH
- DNA nádorová analýza chemie izolace a purifikace MeSH
- fixace tkání MeSH
- kolorektální nádory genetika patologie MeSH
- lidé MeSH
- nádorové buňky kultivované MeSH
- nádory plic genetika patologie MeSH
- RNA nádorová analýza chemie izolace a purifikace MeSH
- vysoce účinné nukleotidové sekvenování metody MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Microsatellite instability (MSI) is present in 15-20% of primary colorectal cancers. MSI status is assessed to detect Lynch syndrome, guide adjuvant chemotherapy, determine prognosis, and use as a companion test for checkpoint blockade inhibitors. Traditionally, MSI status is determined by immunohistochemistry or molecular methods. The Idylla™ MSI Assay is a fully automated molecular method (including automated result interpretation), using seven novel MSI biomarkers (ACVR2A, BTBD7, DIDO1, MRE11, RYR3, SEC31A, SULF2) and not requiring matched normal tissue. In this real-world global study, 44 clinical centers performed Idylla™ testing on a total of 1301 archived colorectal cancer formalin-fixed, paraffin-embedded (FFPE) tissue sections and compared Idylla™ results against available results from routine diagnostic testing in those sites. MSI mutations detected with the Idylla™ MSI Assay were equally distributed over the seven biomarkers, and 84.48% of the MSI-high samples had ≥ 5 mutated biomarkers, while 98.25% of the microsatellite-stable samples had zero mutated biomarkers. The concordance level between the Idylla™ MSI Assay and immunohistochemistry was 96.39% (988/1025); 17/37 discordant samples were found to be concordant when a third method was used. Compared with routine molecular methods, the concordance level was 98.01% (789/805); third-method analysis found concordance for 8/16 discordant samples. The failure rate of the Idylla™ MSI Assay (0.23%; 3/1301) was lower than that of referenced immunohistochemistry (4.37%; 47/1075) or molecular assays (0.86%; 7/812). In conclusion, lower failure rates and high concordance levels were found between the Idylla™ MSI Assay and routine tests.
- MeSH
- fixace tkání * MeSH
- fixativa MeSH
- formaldehyd MeSH
- imunohistochemie * MeSH
- kolorektální nádory chemie genetika patologie MeSH
- laboratorní automatizace MeSH
- lidé MeSH
- mikrosatelitní nestabilita * MeSH
- mutace * MeSH
- mutační analýza DNA * MeSH
- nádorové biomarkery * analýza genetika MeSH
- prediktivní hodnota testů MeSH
- reprodukovatelnost výsledků MeSH
- zalévání tkání do parafínu * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- multicentrická studie MeSH
- srovnávací studie MeSH
