ArticleMolecular pathology : MP2003
Development of molecular methods for the identification of aspergillus and emerging moulds in paraffin wax embedded tissue sections.
Article in Molecular pathology : MP, 2003. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed.
- Invasive Pulmonary Aspergillosis.Journal of fungi (Basel, Switzerland) · 2023Review
- Molecular Techniques for Genus and Species Determination of Fungi From Fresh and Paraffin-Embedded Formalin-Fixed Tissue in the Revised EORTC/MSGERC Definitions of Invasive Fungal Infection.Clinical infectious diseases : an official publication of the Infectious Diseases Society of America · 2021Article
- Panfungal Polymerase Chain Reaction for Identification of Fungal Pathogens in Formalin-Fixed Animal Tissues.Veterinary pathology · 2017Article
- Seven cases of localized invasive sino-orbital aspergillosis.Japanese journal of ophthalmology · 2017Article
- Aspergillus PCR-based investigation of fresh tissue and effusion samples in patients with suspected invasive Aspergillosis enhances diagnostic capabilities.Journal of clinical microbiology · 2013Article
- Development of an ammonium sulfate DNA extraction method for obtaining amplifiable DNA in a small number of cells and its application to clinical specimens.BioMed research international · 2013Article
- Candida kefyr fungal enteritis following autologous BMT.Bone marrow transplantation · 2012Article
- Targeting ST2L potentiates CpG-mediated therapeutic effects in a chronic fungal asthma model.The American journal of pathology · 2011Article
- Identification of Fusarium species in formalin-fixed and paraffin-embedded sections by in situ hybridization using peptide nucleic acid probes.Journal of clinical microbiology · 2011Article
- Triggering receptor expressed on myeloid cells-1 (TREM-1) modulates immune responses to Aspergillus fumigatus during fungal asthma in mice.Immunological investigations · 2011Article
- Improving molecular detection of fungal DNA in formalin-fixed paraffin-embedded tissues: comparison of five tissue DNA extraction methods using panfungal PCR.Journal of clinical microbiology · 2010Article
- Molecular confirmation of invasive infection caused by Chaetomium globosum.Journal of clinical pathology · 2005Article
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4 authors.
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Abstract
BACKGROUND/
aimsInvasive infection with emerging moulds is increasing in incidence and reliable methods for speciating these organisms in tissue sections need to be developed.
methodsTwo methods for extracting fungal DNA from paraffin wax embedded tissue sections, based on the TaKaRa DEXPAT kit and QIAamp DNA mini kit, were optimised and compared. DNA was amplified by PCR using pan-fungal probes, and detected by Southern blot hybridisation using a high stringency method with a probe specific for Aspergillus fumigatus and A flavus.
resultsThe method based on the TaKaRa DEXPAT kit, with additional steps using lyticase and ethanol precipitation, was superior. Less than 10 conidia were detectable using spiked samples and a positive result was obtained with 100% of clinical samples known to be culture positive for A fumigatus. Other moulds could be identified by using species specific probes or by sequencing PCR products.
conclusionsThe method based on the TaKaRa DEXPAT kit could detect less than 10 conidia/sample. The method allowed accurate identification of A fumigatus and A flavus and other species could be identified using species specific probes or by DNA sequencing. These methods will provide a valuable diagnostic tool for both patient management and future antifungal and epidemiological studies.
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