PMID: 9650962Jul 3, 1998Paper

Utility of random amplified polymorphic DNA PCR and TaqMan automated detection in molecular identification of Aspergillus fumigatus

Journal of Clinical Microbiology
Mary BrandtB P Holloway

Abstract

We developed a method for the identification of Aspergillus fumigatus fungal isolates by using random amplified polymorphic DNA (RAPD) PCR (RAPD-PCR) cloning and the TaqMan LS50B fluorogenic detection system (Perkin-Elmer Corp., Applied Biosystems, Foster City, Calif.). DNA from seven clinically important Aspergillus species was screened by RAPD-PCR to identify section- or species-specific amplicons. With the OPZ19 RAPD primer a 1,264-bp product was amplified from all A. fumigatus strains initially examined but not from other species. A partial DNA sequence of this product was used to design a specific primer pair, which generated a single 864-bp fragment with DNA from 90 of 100 A. fumigatus isolates when a "touchdown" (65-->55 degrees C) annealing protocol was used. The TaqMan system, a fluorogenic assay which uses the 5'-->3' endonuclease activity of Taq DNA polymerase, detected this 864-bp product with DNA from 89 of these 90 A. fumigatus strains; 1 DNA sample generated an indeterminate result. With DNA from three morphologically typical A. fumigatus isolates, six white ("albino") A. fumigatus isolates, and five of six Neosartorya species (non-A. fumigatus members of the section Fumigati), the 864-bp product was amplified di...Continue Reading

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Citations

Jul 20, 2001·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·D BuchheidtR Hehlmann
Nov 23, 2018·Genome Génome / Conseil National De Recherches Canada·Wieland MeyerUNKNOWN ISHAM barcoding of pathogenic fungi working group
Sep 25, 2002·Infection Control and Hospital Epidemiology : the Official Journal of the Society of Hospital Epidemiologists of America·Theresa HahnPhilip L McCarthy
Aug 22, 2007·Medical Mycology·S Arunmozhi BalajeeMary E Brandt
Jul 5, 2002·Clinical Microbiology Reviews·Siew Fah Yeo, Brian Wong
Apr 9, 2001·Molecular and Cellular Probes·P Cruz-PerezL D Stetzenbach
Apr 9, 1999·Clinical Microbiology Reviews·J P Latgé
Jul 6, 2000·Journal of Clinical Microbiology·G E KillgoreF C Tenover
May 15, 1999·Journal of Clinical Microbiology·A PahlA Gessner
Mar 11, 2004·Clinical Microbiology and Infection : the Official Publication of the European Society of Clinical Microbiology and Infectious Diseases·I M Mackay
Apr 23, 2011·BMC Microbiology·Rita SerranoRicardo Araujo

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