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Development and validation of a quantitative PCR assay for diagnosis of scedosporiosis
- Source :
- Journal of clinical microbiology. 46(10)
- Publication Year :
- 2008
-
Abstract
- Scedosporium apiospermum and Scedosporium prolificans are fungal pathogens that can cause severe human infections, including disseminated mycosis in immunocompromised patients. Two real-time PCR (RT-PCR) assays for the diagnosis of these species were developed and validated for the classification of clinical strains and for the detection of DNA in clinical samples by use of a murine model of invasive infection. A total of 14 clinical strains and 141 samples, including blood, serum, and lung samples from infected CD1 mice, were analyzed. Each RT-PCR methodology used a species-specific molecular beacon probe targeting a highly conserved region of the fungal ribosomal DNA gene. Results showed 100% specificity and a detection limit of 10 fg of DNA for both assays. The sensitivities for the S. prolificans -specific PCR assay were 100% for cultured clinical strains, 95.5% for lung tissues, 85% for serum, and 83.3% for blood. For S. apiospermum , the sensitivities were 100% for clinical strains and 97.2%, 81.8%, and 54.5% for lung tissues, serum, and blood, respectively. Both techniques can be useful for clinical diagnosis, and further studies are warranted.
- Subjects :
- Microbiology (medical)
Male
Mycology
Polymerase Chain Reaction
Sensitivity and Specificity
Microbiology
law.invention
Scedosporium
Mice
law
medicine
Animals
Humans
DNA, Fungal
Ribosomal DNA
Lung
Polymerase chain reaction
Fluorescent Dyes
Scedosporium prolificans
biology
Fungal genetics
Scedosporium apiospermum
biology.organism_classification
Survival Analysis
medicine.anatomical_structure
Real-time polymerase chain reaction
Blood
Mycoses
Oligonucleotide Probes
Subjects
Details
- ISSN :
- 1098660X
- Volume :
- 46
- Issue :
- 10
- Database :
- OpenAIRE
- Journal :
- Journal of clinical microbiology
- Accession number :
- edsair.doi.dedup.....6f7b751b384bfd3a94b086db01971b90