ArticleJournal of clinical microbiology2000
Development of a PCR-based line probe assay for identification of fungal pathogens.
Article in Journal of clinical microbiology, 2000. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 27 papers.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
27 citing papers in PubMed.
- Long-Reads-Based Metagenomics in Clinical Diagnosis With a Special Focus on Fungal Infections.Frontiers in microbiology · 2021Review
- Development of a Cost-Effective Line Probe Assay for Rapid Detection and Differentiation ofReports of biochemistry & molecular biology · 2020Article
- The Role of PCR in the Diagnosis of Candida Vulvovaginitis-a New Gold Standard?Current infectious disease reports · 2015Article
- Candida identification: a journey from conventional to molecular methods in medical mycology.World journal of microbiology & biotechnology · 2014Review
- Enzymatic treatment of specimens before DNA extraction directly influences molecular detection of infectious agents.PloS one · 2014Article
- Rapid identification of allergenic and pathogenic molds in environmental air by an oligonucleotide array.BMC infectious diseases · 2011Article
- Identification of fungal species by fragment length analysis of the internally transcribed spacer 2 region.European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology · 2009Article
- Evaluation of novel broad-range real-time PCR assay for rapid detection of human pathogenic fungi in various clinical specimens.Journal of clinical microbiology · 2008Article
- DNA microarray based on arrayed-primer extension technique for identification of pathogenic fungi responsible for invasive and superficial mycoses.Journal of clinical microbiology · 2008Article
- Analysis of fungal flora in indoor dust by ribosomal DNA sequence analysis, quantitative PCR, and culture.Applied and environmental microbiology · 2008Article
- Reverse line blot hybridization assay for identification of medically important fungi from culture and clinical specimens.Journal of clinical microbiology · 2007Article
- High-throughput identification of clinical pathogenic fungi by hybridization to an oligonucleotide microarray.Journal of clinical microbiology · 2006Article
- Identification of medically important yeast species by sequence analysis of the internal transcribed spacer regions.Journal of clinical microbiology · 2006Article
- Simultaneous detection and identification of Candida, Aspergillus, and Cryptococcus species by reverse line blot hybridization.Journal of clinical microbiology · 2006Article
- Diagnosis of candidemia by polymerase chain reaction and blood culture: prospective study in a high-risk population and identification of variables associated with development of candidemia.European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology · 2005Article
- Development of a DNA microarray for detection and identification of fungal pathogens involved in invasive mycoses.Journal of clinical microbiology · 2005Article
- Identification of medically important molds by an oligonucleotide array.Journal of clinical microbiology · 2005Article
- Assessment of ribosomal large-subunit D1-D2, internal transcribed spacer 1, and internal transcribed spacer 2 regions as targets for molecular identification of medically important Aspergillus species.Journal of clinical microbiology · 2005Article
- An interlaboratory comparison of ITS2-PCR for the identification of yeasts, using the ABI Prism 310 and CEQ8000 capillary electrophoresis systems.BMC microbiology · 2005Article
- Amplification of coccidioidal DNA in clinical specimens by PCR.Journal of clinical microbiology · 2004Article
Corrections and comments
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Authors and funding
7 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
We report on a reverse-hybridization line probe assay (LiPA) which when combined with PCR amplification detects and identifies clinically significant fungal pathogens including Candida, Aspergillus, and Cryptococcus species. DNA probes have been designed from the internal transcribed-spacer (ITS) regions of Candida albicans, Candida parapsilosis, Candida glabrata, Candida tropicalis, Candida krusei, Candida dubliniensis, Cryptococcus neoformans, Aspergillus fumigatus, Aspergillus versicolor, Aspergillus nidulans and Aspergillus flavus. The probes were incorporated into a LiPA for detection of biotinylated ITS PCR products, and the specificity of the probes was evaluated. We established LiPA detection limits for ITS 1 and for full ITS amplicons for genomic DNA from C. albicans, A. fumigatus, and C. neoformans. Further evaluation of the LiPA was carried out on clinical fungal isolates. One hundred twenty-seven isolates consisting of dimorphic yeasts and dermatophytic and filamentous fungi were tested by the LiPA, which correctly identified 77 dimorphic yeasts and 23 of the filamentous isolates; the remaining 27 isolates represented species of fungi for which probes were not included in the LiPA. The fungal-PCR-LiPA technology was applied to blood samples inoculated with Candida cells which were pretreated by minibead beating to mechanically disrupt the cells, with the DNA extracted by either a previously described guanidium thiocyanate-silica method or the commercially available QIAmp tissue kit. PCR amplification of the extracted DNA and subsequent DNA probe hybridization in the LiPA assay yielded detection limits of 2 to 10 cells/ml. An internal standard control was included in the PCR amplification to monitor for PCR inhibition. This fungal PCR-LiPA assay is robust and sensitive and can easily be integrated into a clinical-testing laboratory with the potential for same-day diagnosis of fungal infection.
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