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Utilization of the internal transcribed spacer regions as molecular targets to detect and identify human fungal
P C Iwen1, S H Hinrichs, M E Rupp
1Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha 68198-6495, USA. piwen@unmc.edu
Abstract:
Advances in molecular technology show great potential for the rapid detection and identification of fungi for medical, scientific and commercial purposes. Numerous targets within the fungal genome have been evaluated, with much of the current work using sequence areas within the ribosomal DNA (rDNA) gene complex. This section of the genome includes the 18S, 5.8S and 28S genes which code for ribosomal RNA (rRNA) and which have a relatively conserved nucleotide sequence among fungi. It also includes the variable DNA sequence areas of the intervening internal transcribed spacer (ITS) regions called ITS1 and ITS2. Although not translated into proteins, the ITS coding regions have a critical role in the development of functional rRNA, with sequence variations among species showing promise as signature regions for molecular assays. This review of the current literature was conducted to evaluate clinical approaches for using the fungal ITS regions as molecular targets. Multiple applications using the fungal ITS sequences are summarized here including those for culture identification, phylogenetic research, direct detection from clinical specimens or the environment, and molecular typing for epidemiological investigations. The breadth of applications shows that ITS regions have great potential as targets in molecular-based assays for the characterization and identification of fungi. Development of rapid and accurate amplification-based ITS assays to diagnose invasive fungal infections could potentially impact care and improve outcome for affected patients.
Insights
Molecular technology enables rapid fungal detection using internal transcribed spacer (ITS) regions. These fungal DNA targets show promise for diagnosing infections and advancing research.
Area of Science:
- Mycology
- Molecular Biology
- Genetics
Background:
- Molecular technologies offer rapid fungal detection and identification for various applications.
- The fungal ribosomal DNA (rDNA) gene complex, including conserved rRNA genes and variable internal transcribed spacer (ITS) regions (ITS1 and ITS2), is a key target.
- ITS regions, crucial for rRNA function, exhibit species-specific variations valuable for molecular assays.
Purpose of the Study:
- To review and evaluate clinical approaches utilizing fungal ITS regions as molecular targets.
- To summarize current applications of fungal ITS sequences in molecular assays.
- To assess the potential of ITS regions for fungal characterization and identification.
Main Methods:
- Literature review of current scientific publications.
- Evaluation of studies employing fungal ITS sequences for molecular assays.
- Analysis of applications in culture identification, phylogenetic research, direct detection, and epidemiological typing.
Main Results:
- Fungal ITS regions are versatile targets with broad applications in molecular assays.
- ITS sequences are effective for fungal culture identification, phylogenetic analysis, and direct detection from clinical or environmental samples.
- Molecular typing using ITS regions aids in epidemiological investigations of fungal infections.
Conclusions:
- Fungal ITS regions demonstrate significant potential as targets for molecular-based fungal identification and characterization.
- Development of rapid, amplification-based ITS assays can improve the diagnosis of invasive fungal infections.
- Accurate ITS assays hold the potential to positively impact patient care and outcomes for invasive fungal infections.