Dataset Information

Diagnostic information Profiling and Evaluation of Causative Fungi of Fungal Keratitis Using High-throughput Internal Transcribed Spacer Sequencing.

ABSTRACT: Early and accurate diagnosis is essential for the targeted management of fungal keratitis (FK), which is one of the major blinding eye diseases worldwide. To elucidate the diagnostic information of high-throughput internal transcribed spacer (ITS) sequencing for identifying causative fungi of FK, 38 patients who were highly suspected of having FK were included in this research. In vivo confocal microscopy, potassium hydroxide smear, and fungal culture were performed to diagnose FK. Culture and ITS sequencing were used to identify causative fungi. We hypothesized that the dominant genus was the result of pathogen identification by ITS sequencing. Thirty-five patients were eventually diagnosed with FK, with fungal pathogens found by confocal microscopy in 27 patients (77.14%), by smear examination in 27 patients (77.14%), by culture in 25 patients (71.43%), and by ITS sequencing in 26 patients (74.29%). Eight causative fungal genera were determined by ITS sequencing, while five causative fungal genera were identified based on the morphology of the cultured pathogens. The results of ITS sequencing and culture were coincident in 10 patients with FK (28.57%). It is concluded that ITS sequencing, to some extent, challenged fungal culture and might be an optional complement in identifying fungal pathogens in corneas.

SUBMITTER: Ren Z

PROVIDER: S-EPMC6997210 | biostudies-literature | 2020 Feb

REPOSITORIES: biostudies-literature

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Diagnostic information Profiling and Evaluation of Causative Fungi of Fungal Keratitis Using High-throughput Internal Transcribed Spacer Sequencing.

Ren Zhichao Z Liu Qing Q Wang Yuqian Y Dong Yanling Y Huang Yusen Y

Scientific reports 20200203 1

Early and accurate diagnosis is essential for the targeted management of fungal keratitis (FK), which is one of the major blinding eye diseases worldwide. To elucidate the diagnostic information of high-throughput internal transcribed spacer (ITS) sequencing for identifying causative fungi of FK, 38 patients who were highly suspected of having FK were included in this research. In vivo confocal microscopy, potassium hydroxide smear, and fungal culture were performed to diagnose FK. Culture and I ...[more]

PMID: 32015395

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Project description:The health, growth, and fitness of boreal forest trees are impacted and improved by their associated microbiomes. Microbial gene expression and functional activity can be assayed with RNA sequencing (RNA-Seq) data from host samples. In contrast, phylogenetic marker gene amplicon sequencing data are used to assess taxonomic composition and community structure of the microbiome. Few studies have considered how much of this structural and taxonomic information is included in transcriptomic data from matched samples. Here, we described fungal communities using both host-derived RNA-Seq and fungal ITS1 DNA amplicon sequencing to compare the outcomes between the methods. We used a panel of root and needle samples from the coniferous tree species Picea abies (Norway spruce) growing in untreated (nutrient-deficient) and nutrient-enriched plots at the Flakaliden forest research site in boreal northern Sweden. We show that the relationship between samples and alpha and beta diversity indicated by the fungal transcriptome is in agreement with that generated by the ITS data, while also identifying a lack of taxonomic overlap due to limitations imposed by current database coverage. Furthermore, we demonstrate how metatranscriptomics data additionally provide biologically informative functional insights. At the community level, there were changes in starch and sucrose metabolism, biosynthesis of amino acids, and pentose and glucuronate interconversions, while processing of organic macromolecules, including aromatic and heterocyclic compounds, was enriched in transcripts assigned to the genus CortinariusIMPORTANCE A deeper understanding of microbial communities associated with plants is revealing their importance for plant health and productivity. RNA extracted from plant field samples represents the host and other organisms present. Typically, gene expression studies focus on the plant component or, in a limited number of studies, expression in one or more associated organisms. However, metatranscriptomic data are rarely used for taxonomic profiling, which is currently performed using amplicon approaches. We created an assembly-based, reproducible, and hardware-agnostic workflow to taxonomically and functionally annotate fungal RNA-Seq data obtained from Norway spruce roots, which we compared to matching ITS amplicon sequencing data. While we identified some limitations and caveats, we show that functional, taxonomic, and compositional insights can all be obtained from RNA-Seq data. These findings highlight the potential of metatranscriptomics to advance our understanding of interaction, response, and effect between host plants and their associated microbial communities.

Dataset Information

Diagnostic information Profiling and Evaluation of Causative Fungi of Fungal Keratitis Using High-throughput Internal Transcribed Spacer Sequencing.

Publications

Diagnostic information Profiling and Evaluation of Causative Fungi of Fungal Keratitis Using High-throughput Internal Transcribed Spacer Sequencing.

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