Utilization of the internal transcribed spacer regions as molecular targets to detect and identify human fungal pathogens.
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.
Iwen
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Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha 68198-6495, USA. piwen@unmc.edu
Hinrichs
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Rupp
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D021903:DNA, Ribosomal Spacer / Q000737:chemistry / Q000235:genetics*; D005658:Fungi / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D015345:Oligonucleotide Probes; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D013045:Species Specificity
D021903:DNA, Ribosomal Spacer; D015345:Oligonucleotide Probes