PCR-RFLP detection and species identification of fungal pathogens in patients with febrile neutropenia
Jazyk angličtina Země Anglie, Velká Británie Médium print
Typ dokumentu časopisecké články, práce podpořená grantem
PubMed
14686984
DOI
10.1111/j.1469-0691.2003.00719.x
PII: S1198-743X(14)63226-2
Knihovny.cz E-zdroje
- MeSH
- dítě MeSH
- DNA fungální chemie genetika MeSH
- horečka mikrobiologie MeSH
- hostitel s imunodeficiencí MeSH
- houby genetika izolace a purifikace MeSH
- kojenec MeSH
- lidé MeSH
- mezerníky ribozomální DNA chemie genetika MeSH
- mladiství MeSH
- mykózy mikrobiologie MeSH
- neutropenie mikrobiologie MeSH
- polymerázová řetězová reakce metody MeSH
- polymorfismus délky restrikčních fragmentů MeSH
- předškolní dítě MeSH
- RNA ribozomální 5.8S chemie genetika MeSH
- RNA ribozomální chemie genetika MeSH
- senzitivita a specificita MeSH
- Check Tag
- dítě MeSH
- kojenec MeSH
- lidé MeSH
- mladiství MeSH
- mužské pohlaví MeSH
- předškolní dítě MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- DNA fungální MeSH
- mezerníky ribozomální DNA MeSH
- RNA ribozomální 5.8S MeSH
- RNA ribozomální MeSH
- RNA, ribosomal, 25S MeSH Prohlížeč
OBJECTIVE: To assess the usefulness of polymerase chain reaction (PCR) assays in the diagnosis of fungal infections in immunocompromised patients. METHODS: A rapid and sensitive PCR-based assay for the detection and identification of fungal pathogens was designed and applicability of this method was investigated in a group of children with cancer and febrile neutropenia (FN). RESULTS: The ITS2 sequences and adjacent regions of 40 fungal pathogens were analyzed and primers for detection of all analyzed fungal species were designed. Amplification product length polymorphism (APLP) and restriction fragment length polymorphism (RFLP) generated genus- or species-specific patterns. The sensitivity of the method was approximately three cells of Candida albicans per 1 mL of blood. The results were available within 8 h after sample collection. The method was tested on 53 blood samples and one lung biopsy sample from 24 children with cancer and febrile neutropenia (FN). The PCR assay detected fungal DNA in 25 clinical samples from ten patients. Blood cultures were positive in only five samples, while another two blood-culture negative patients had positive cultures from throat swabs. The remaining 14 patients were both culture- and PCR-negative. Culture-isolated strains matched completely those obtained by PCR-APLP-RFLP identification. The identity of fungal species was confirmed by direct sequencing of amplified products. CONCLUSION: Our results suggest that PCR-APLP-RFLP assays can be useful in the diagnosis of fungal infections in immunocompromised patients.
Citace poskytuje Crossref.org
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