scholarly journals Evaluating a semi‐nested PCR to support histopathology reports of fungal rhinosinusitis in formalin‐fixed paraffin‐embedded tissue samples

2022 ◽  
Author(s):  
Mohammad Javad Ashraf ◽  
Mohammad Kord ◽  
Hamid Morovati ◽  
Saham Ansari ◽  
Golsa Shekarkhar ◽  
...  
Keyword(s):  
Nested Pcr ◽  
Tissue Samples ◽  
Fungal Rhinosinusitis ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Formalin Fixed

scholarly journals Evaluating A Semi-Nested PCR to Support Histopathology Reports of Fungal Rhinosinusitis in Formalin-Fixed Paraffin-Embedded Tissue Samples.

2021 ◽  
Author(s):  
Mohammad Javad Ashraf ◽  
Mohammad Kord ◽  
Hamid Morovati ◽  
Saham Ansari ◽  
Golsa Shekarkhar ◽  
...  
Keyword(s):  
Nested Pcr ◽  
Predictive Value ◽  
Globin Gene ◽  
Sinus Surgery ◽  
Invasive Infection ◽  
Internal Transcribed Spacer Region ◽  
Fungal Rhinosinusitis ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Formalin Fixed

Abstract Background Fungal rhinosinusitis (FRS) encompasses a various spectrum of disease, which vary in clinical presentation, histologic features, and biological significance. FRS is commonly classified in two categories, i.e., invasive and non-invasive infection. Histopathology is the “gold standard” diagnostic method, but it is not able to determine the genus and species. Almost more than 50% of the histopathologically proven cases, the culture elicited no reliable results. This study was an attempt to evaluate the diagnostic efficiency of semi-nested polymerase chain reaction (PCR) to confirmation of pathology reports obtained from formalin-fixed paraffin-embedded (FFPE) functional endoscopic sinus surgery (FESS) in FRS patients. Methods One hundred ten specimens were subjected to DNA extraction and histopathology examination. The amplification of β-globin gene by conventional PCR was used for confirming the quality of extracted DNA. The semi-nested PCR was performed using ITS 1 and ITS4 primers during two steps. In order to identification of causative agents, sequencing of the internal transcribed spacer region (ITS1-5.8S-ITS2) was performed on PCR products. Results Sixty-four out of 110 samples were positive by histopathology evidence, of which 56 samples (87.5%) were positive by PCR and 8 (12.5%) samples were negative. Out of 46 negative samples by histopathological methods, 41 samples (89.1%) were negative in PCR while 5 (10.9%) were positive. Sensitivity, specificity, positive predictive value, and negative predictive value of the semi-nested PCR method were reported 87.5%, 89.2%, 92.7%, and 85.2%, respectively. Aspergillus flavus, Rhizopus oryzae, Lichtheimia corymbifera and Candida albicans have identified as common fungal species. Conclusion Based on staining and direct examination that are time-consuming, applying molecular methods might be an appropriate choice for rapid diagnosis and support the consequences of histopathology examinations. We suggest that fresh biopsy specimens elicit more reliable results in comparison with FFPE samples. It is recommended that semi-nested PCR assays be performed in a single tube, which showed less prone to contamination when compared with assays that were carried out in two stages and in separate tubes. Trial registration: Not applicable.

scholarly journals The Dutch National TissueArchive Portal enables efficient, consistent, and transparent procurement of diagnostic tissue samples for scientific use

2021 ◽  
Author(s):  
Robin Verjans ◽  
Annette H. Bruggink ◽  
Robby Kibbelaar ◽  
Jos Bart ◽  
Aletta Debernardi ◽  
...  
Keyword(s):  
The Netherlands ◽  
Tissue Sample ◽  
Ffpe Tissue ◽  
Tissue Samples ◽  
Internet Portal ◽  
Formalin Fixed Paraffin ◽  
Practical Support ◽  
Formalin Fixed Paraffin Embedded ◽  
The Rich ◽  
Formalin Fixed

AbstractBiobanks play a crucial role in enabling biomedical research by facilitating scientific use of valuable human biomaterials. The PALGA foundation—a nationwide network and registry of histo- and cytopathology in the Netherlands—was established to promote the provision of data within and between pathology departments, and to make the resulting knowledge available for healthcare. Apart from the pathology data, we aimed to utilize PALGA’s nationwide network to find and access the rich wealth of Formalin-Fixed Paraffin-Embedded (FFPE) tissue samples for scientific use.  We implemented the Dutch National TissueArchive Portal (DNTP) to utilize PALGA’s nationwide network for requesting FFPE tissue samples. The DNTP consists of (1) a centrally organized internet portal to improve the assessing, processing, harmonization, and monitoring of the procurement process, while (2) dedicated HUB-employees provide practical support at peripheral pathology departments. Since incorporation of the DNTP, both the number of filed requests for FFPE tissue samples and the amount of HUB-mediated support increased 55 and 29% respectively. In line, the sample procurement duration time decreased significantly (− 47%). These findings indicate that implementation of the DNTP improved the frequency, efficiency, and transparency of FFPE tissue sample procurement for research in the Netherlands. To conclude, the need for biological resources is growing persistently to enable precision medicine. Here, we access PALGA’s national, pathology network by implementation of the DNTP to allow for efficient, consistent, and transparent exchange of FFPE tissue samples for research across the Netherlands.

Microsatellite instability evaluation: which test to use for endometrial cancer?

2021 ◽  
pp. jclinpath-2021-207723
Author(s):  
Paola Rafaniello-Raviele ◽  
Ilaria Betella ◽  
Alessandra Rappa ◽  
Davide Vacirca ◽  
Gianluca Tolva ◽  
...  
Keyword(s):  
Endometrial Cancer ◽  
Microsatellite Instability ◽  
Automated System ◽  
Tissue Samples ◽  
Valuable Alternative ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Formalin Fixed ◽  
Mmr Proteins

AimsAnalysis of microsatellite instability (MSI) is strongly recommended in endometrial cancer (EC) and colorectal cancer to screen for Lynch syndrome, to predict prognosis and to determine optimal treatment and follow-up. In a large monoinstitutional series of ECs, we evaluated the reliability and accuracy of Idylla assay, a rapid, fully automated system to detect MSI, and we compared its performance with two routine reference methods.MethodsWe evaluated MSI status in 174 formalin-fixed, paraffin-embedded EC tissue samples using immunohistochemistry (IHC) for mismatch repair (MMR) proteins and Idylla assay. Samples with discordant or equivocal results were analysed with a third technique, the Promega MSI kit.ResultsIdylla MSI assay and IHC were highly concordant (overall agreement: 154/170=90.59%, 95% CI 85.26% to 94.12%). However, in four samples, MMR-IHC staining was equivocal; moreover, 16 cases showed discordant results, that is, MMR deficient using IHC and microsatellite stable using Idylla. These 20 samples were reanalysed using the MSI-Promega kit, which showed the same results of Idylla assay in 18/20 cases (overall agreement: 90%, 95% CI 69.90% to 97.21%).ConclusionsOur results suggest that IHC is an efficient method to determine MMR status in ECs. However, the Idylla MSI assay is a rapid and reliable tool to define MSI status, and it could represent a valuable alternative to conventional MSI-PCR methods.

scholarly journals Evaluation of Formalin-fixed Paraffin-embedded Tissue Samples Diagnosed by Histopathology as Echinococcus in Çorum

2021 ◽  
Vol 45 (4) ◽  
pp. 262-267
Author(s):  
Yılmaz Baş ◽  
Yunus Emre Beyhan ◽  
Havva Hande Keser Şahin ◽  
Tuğba Özçerezci ◽  
Dursun Karasartova ◽  
...  
Keyword(s):  
Tissue Samples ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Formalin Fixed

scholarly journals Evaluation of 3 molecular-based assays for microsatellite instability detection in formalin-fixed tissues of patients with endometrial and colorectal cancers

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Pauline Gilson ◽  
Julien Levy ◽  
Marie Rouyer ◽  
Jessica Demange ◽  
Marie Husson ◽  
...  
Keyword(s):  
Microsatellite Instability ◽  
Sensitivity And Specificity ◽  
Tissue Samples ◽  
Fast Screening ◽  
Formalin Fixed Paraffin ◽  
Golden Standard ◽  
Formalin Fixed Paraffin Embedded ◽  
Analysis System ◽  
Msi Analysis ◽  
Formalin Fixed

Abstract Microsatellite instability (MSI) status is routinely assessed in patients with colorectal and endometrial cancers as it contributes to Lynch syndrome initial screening, tumour prognosis and selecting patients for immunotherapy. Currently, standard reference methods recommended for MSI/dMMR (deficient MisMatch Repair) testing consist of immunohistochemistry and pentaplex PCR-based assays, however, novel molecular-based techniques are emerging. Here, we aimed to evaluate the performance of a custom capture-based NGS method and the Bio-Rad ddPCR and Idylla approaches for the determination of MSI status for theranostic purposes in 30 formalin-fixed paraffin embedded (FFPE) tissue samples from patients with endometrial (n = 15) and colorectal (n = 15) cancers. All samples were previously characterised using IHC and Promega MSI Analysis System and these assays set as golden standard. Overall agreement, sensitivity and specificity of our custom-built NGS panel were 93.30%, 93.75% and 92.86% respectively. Overall agreement, sensitivity and specificity were 100% with the Idylla MSI system. The Bio-Rad ddPCR MSI assay showed a 100% concordance, sensitivity and specificity. The custom capture-based NGS, Bio-Rad ddPCR and Idylla approaches represent viable and complementary options to IHC and Promega MSI Analysis System for the detection of MSI. Bio-Rad ddPCR and Idylla MSI assays accounts for easy and fast screening assays while the NGS approach offers the advantages to simultaneously detect MSI and clinically relevant genomic alterations.

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