MARA M DIAZ
Medical Practice at 11, Miami, FL
License number
Florida 305563
Issued Date
Aug 30, 2002
Expiration Date
Dec 1, 2004
Category
Health Care
Type
Emergency Medical Technician
Address
Address
751 SW 11Th St APT 1, Miami, FL 33129
Professional information
High Through-Put Detection Of Pathogenic Yeasts In The Genus Trichosporon
US Patent:
2006021, Sep 28, 2006
Filed:
May 23, 2005
Appl. No.:
11/134619
Inventors:
Mara Diaz - Key Biscayne FL, US
Jack Fell - Miami FL, US
Jack Fell - Miami FL, US
Assignee:
University of Miami - Miami FL
International Classification:
C12Q 1/68, C07H 21/04
US Classification:
435006000, 536024100
Abstract:
The emergence of opportunistic and antifungal resistant strains has given rise to an urgent need for a rapid and accurate method for the detection of fungal pathogens. In this application, we demonstrate the detection of medically important fungal pathogens at the species level. The present method, which is based on a nucleotide hybridization assay, consists of a combination of different sets of fluorescent beads covalently bound to species specific capture probes. Upon hybridization, the beads bearing the target amplicons are classified by their spectral addresses with a 635 nm laser. Quantitation of the hybridized biotinylated amplicon is based on the fluorescent detection with a 532 nm laser. Using this technology we designed and tested various multiplex formats, the performance of forty eight species specific and group specific capture probes designed from sequence analysis in the D1/D2 region of ribosomal DNA, internal transcribed spacer regions (ITS), and intergenic spacer region (IGS). Species-specific biotinylated amplicons (>600 bp) were generated with three sets of primers to yield fragments from the three regions. The developed assay was specific and relatively fast, as it discriminated species differing by one nucleotide and required less than 50 min following amplification to process a 96 well plate with the capability to detect up to 100 species per well. The sensitivity of the assay allowed the detection as low as 10genome molecules in PCR reactions and 10to 10molecules of biotinylated amplification product. This technology provided a rapid means of detection of species and had the flexibility to identify species in a multiplex format by combining different sets of beads. The assay can be expanded to include all known pathogenic fungal species.
Rapid Identification Of The Varieties And Genotypes Of Cryptococcus Neoformans Species Complex Using A High-Throughput Flow Cytometer
US Patent:
2006027, Dec 7, 2006
Filed:
May 17, 2006
Appl. No.:
11/435291
Inventors:
Mara Diaz - Key Biscayne FL, US
Jack Fell - Miami FL, US
Jack Fell - Miami FL, US
Assignee:
University of Miami - Miami FL
International Classification:
C12Q 1/68, C07H 21/04, C12M 1/34
US Classification:
435006000, 435287200, 536024100
Abstract:
Nucleic acid probes and molecular method to identify the varieties and genotypic groups within species complex. The method employs a flow cytometer with a dual laser system that allows the simultaneous detection of different target sequences in a multiplex and high-throughput format. The assay uses a liquid suspension hybridization format with specific oligonucleotide probes that are covalently bound to the surface of fluorescent color-coded microspheres. Biotinylated target amplicons, which hybridized to their complementary probe sequences, are quantified by the addition of the conjugate, streptavidin-R-phycoerythrin. The assay is specific and sensitive, and allows discrimination of 1 bp mismatch with no apparent cross-reactivity and is capable of detecting 10to 10genome copies. The assay can be used directly with yeast cells or isolated DNA, can be undertaken in less than one hour following PCR amplification and permits identification of species in a multiplex format. In addition, to multiplex capability, the assay allows simultaneous detection of target sequences in a single reaction.