Histopathology of fungal disease

2017 ◽  
Author(s):  
Sebastian B. Lucas
Keyword(s):  
Fungal Infections ◽  
Critical Role ◽  
Species Level ◽  
Molecular Diagnostic ◽  
Species Discrimination ◽  
Autopsy Material ◽  
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Histopathology has a critical role in the diagnosis of fungal infections. Often it is the first or only sample of a lesion. A rapid, confident diagnosis can significantly affect patient management. However, the morphologies of yeast and hyphae are not necessarily diagnostic at the genus or species level, and the experience of histopathologists is variable. A primary decision is whether the lesion is fungal or another infection or not infectious at all, and the next is whether the fungus is a yeast or a hyphal (mould) infection. Further histopathological genus and species discrimination can be made in many cases, but not all. Increasingly, molecular diagnostic DNA technology works effectively on formalin-fixed paraffin-embedded biopsy/autopsy material, and such information can be added to the multidisciplinary input for an optimal diagnosis.

scholarly journals Identification and Validation of New Cancer Stem Cell-Related Genes and Their Regulatory microRNAs in Colorectal Cancerogenesis

Biomedicines ◽  
2021 ◽  
Vol 9 (2) ◽  
pp. 179
Author(s):  
Kristian Urh ◽  
Margareta Žlajpah ◽  
Nina Zidar ◽  
Emanuela Boštjančič
Keyword(s):  
Experimental Validation ◽  
Target Gene ◽  
Bioinformatics Analysis ◽  
Early Carcinoma ◽  
Significant Progress ◽  
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Node Metastases ◽  
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Significant progress has been made in the last decade in our understanding of the pathogenetic mechanisms of colorectal cancer (CRC). Cancer stem cells (CSC) have gained much attention and are now believed to play a crucial role in the pathogenesis of various cancers, including CRC. In the current study, we validated gene expression of four genes related to CSC, L1TD1, SLITRK6, ST6GALNAC1 and TCEA3, identified in a previous bioinformatics analysis. Using bioinformatics, potential miRNA-target gene correlations were prioritized. In total, 70 formalin-fixed paraffin-embedded biopsy samples from 47 patients with adenoma, adenoma with early carcinoma and CRC without and with lymph node metastases were included. The expression of selected genes and microRNAs (miRNAs) was evaluated using quantitative PCR. Differential expression of all investigated genes and four of six prioritized miRNAs (hsa-miR-199a-3p, hsa-miR-335-5p, hsa-miR-425-5p, hsa-miR-1225-3p, hsa-miR-1233-3p and hsa-miR-1303) was found in at least one group of CRC cancerogenesis. L1TD1, SLITRK6, miR-1233-3p and miR-1225-3p were correlated to the level of malignancy. A negative correlation between miR-199a-3p and its predicted target SLITRK6 was observed, showing potential for further experimental validation in CRC. Our results provide further evidence that CSC-related genes and their regulatory miRNAs are involved in CRC development and progression and suggest that some them, particularly miR-199a-3p and its SLITRK6 target gene, are promising for further validation in CRC.

scholarly journals Immunomycology: rapid and specific immunocytochemical identification of fungi in formalin-fixed, paraffin-embedded material.

1993 ◽  
Vol 41 (8) ◽  
pp. 1217-1221 ◽  
Author(s):  
J A Reed ◽  
B A Hemann ◽  
J L Alexander ◽  
D J Brigati
Keyword(s):  
Complement Fixation ◽  
Fungal Infections ◽  
Periodic Acid ◽  
Cross Reactivity ◽  
Periodic Acid Schiff ◽  
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Identification Of Fungi ◽  
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We report the rapid (less than 1 hr), immunocytochemical identification of various fungi in formalin-fixed, paraffin-embedded tissues using antisera originally developed for use in immunodiffusion assays. Primary antisera directed towards fungal genera including Aspergillus, Blastomyces, Candida, Coccidioides, Cryptococcus, Histoplasma, and Sporothrix were examined. The specificity of each antiserum was evaluated by the presence or absence of crossreactivity with other morphologically similar fungi in both paraffin-embedded pure fungal cultures and tissues with culture-confirmed fungal infections. Each antiserum reacted strongly with the fungus to which it had been raised, whether examined in pure culture or infected tissues. The antisera raised against Candida, Cryptococcus, and Sporothrix did not exhibit cross-reactivity with any other fungus tested. However, the antisera raised to Aspergillus, Blastomyces, Coccidioides, and Histoplasma demonstrated significant crossreactivity with other genera of fungi, thus precluding their routine use in diagnostic immunocytochemistry. The results indicate that immunocytochemistry may provide an important adjunct to other methods, such as immunodiffusion or complement fixation assays and histochemical stains such as the Grocott methenamine silver or periodic acid-Schiff, when attempts are made to specifically identify certain fungi in formalin-fixed, paraffin-embedded tissues before mycology culture results are available.

Validating a fully automated real-time PCR-based system for use in the molecular diagnostic analysis of colorectal carcinoma: a comparison with NGS and IHC

2017 ◽  
Vol 70 (7) ◽  
pp. 610-614 ◽  
Author(s):  
Richard Colling ◽  
Lai Mun Wang ◽  
Elizabeth Soilleux
Keyword(s):  
Colorectal Carcinoma ◽  
Digital Pcr ◽  
Molecular Diagnostic ◽  
Molecular Testing ◽  
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Polymerase Chain ◽  
Next Generation Sequencing Ngs ◽  
Generation Sequencing ◽  
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BackgroundMolecular testing is increasingly needed in colorectal carcinoma (CRC) and the current clinically relevant mutations are in BRAF, KRAS and NRAS. This study aimed to further validate a new alternative polymerase chain reaction (PCR) platform (Idylla, Biocartis) against existing next-generation sequencing (NGS) and immunohistochemistry (IHC) assays.Methods56 Idylla tests were performed on 43 CRC cases, in a total of 74 comparisons against an NGS panel (Ion Torrent) and the VE1 (anti-BRAF) antibody IHC. Discrepant cases were also compared with either conventional (Cobas) or droplet digital PCR (Bio-Rad).ResultsIdylla showed an overall concordance of 100% (95% CI 93% to 100%) with comparator molecular testing and indications were that Idylla is likely to be more sensitive than routine NGS. BRAF IHC showed 90% concordance with NGS (95% CI 70% to 97%).ConclusionsThis study validates Idylla in formalin-fixed, paraffin-embedded CRC tissue. BRAF IHC, however, is an unreliable substitute for molecular testing in CRC.

scholarly journals Detection of Histoplasma DNA from Tissue Blocks by a Specific and a Broad-Range Real-Time PCR: Tools to Elucidate the Epidemiology of Histoplasmosis

2020 ◽  
Vol 6 (4) ◽  
pp. 319
Author(s):  
Dunja Wilmes ◽  
Ilka McCormick-Smith ◽  
Charlotte Lempp ◽  
Ursula Mayer ◽  
Arik Bernard Schulze ◽  
...  
Keyword(s):  
Diagnostic Tests ◽  
Fungal Infections ◽  
Ffpe Tissue ◽  
Clinical Sensitivity ◽  
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Three Samples ◽  
Ffpe Samples ◽  
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Tissue Specimens ◽  
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Lack of sensitive diagnostic tests impairs the understanding of the epidemiology of histoplasmosis, a disease whose burden is estimated to be largely underrated. Broad-range PCRs have been applied to identify fungal agents from pathology blocks, but sensitivity is variable. In this study, we compared the results of a specific Histoplasma qPCR (H. qPCR) with the results of a broad-range qPCR (28S qPCR) on formalin-fixed, paraffin-embedded (FFPE) tissue specimens from patients with proven fungal infections (n = 67), histologically suggestive of histoplasmosis (n = 36) and other mycoses (n = 31). The clinical sensitivity for histoplasmosis of the H. qPCR and the 28S qPCR was 94% and 48.5%, respectively. Samples suggestive for other fungal infections were negative with the H. qPCR. The 28S qPCR did not amplify DNA of Histoplasma in FFPE in these samples, but could amplify DNA of Emergomyces (n = 1) and Paracoccidioides (n = 2) in three samples suggestive for histoplasmosis but negative in the H. qPCR. In conclusion, amplification of Histoplasma DNA from FFPE samples is more sensitive with the H. qPCR than with the 28S qPCR. However, the 28S qPCR identified DNA of other fungi in H. qPCR-negative samples presenting like histoplasmosis, suggesting that the combination of both assays may improve the diagnosis.

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