Methods and compositions for detecting Aspergillus terreus, Aspergillus niger, and mycotoxins
Inventors
Assignees
Mycodart Inc • MEDICAL SERVICE CONSULTATION INTERNATIONAL LLC
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-8956821-B2
Publication Date
2015-02-17
Expiration Date
Abstract
The invention relates to a method of identifying an Aspergillus terreus or an Aspergillus niger fungal species in patient tissue or body fluid and to primers and probes for use in such a method.
Core Innovation
The invention provides diagnostic methods and compositions for identifying an Aspergillus terreus fungal species in a patient tissue or a patient body fluid. The method includes extracting and recovering DNA of the Aspergillus terreus fungal species from the patient tissue or the patient body fluid, amplifying the DNA, hybridizing a probe to the DNA, and specifically identifying the Aspergillus terreus fungal species. The probe consists of SEQ ID NO: 1 for identifying Aspergillus terreus.
The described approach uses probe- and primer-based PCR DNA amplification with specificity testing to select probe/primer designs that uniquely detect the target species while non-selected designs cross-react with other Aspergillus species. In addition to standard amplifying DNA, the disclosure includes real-time PCR and probe formats including fluorescent labeling and bead dyed with a fluorochrome.
The disclosure further includes constraints on the amplified DNA, including using internal transcribed spacer regions of nuclear ribosomal DNA. It also describes that the method can be applied to a patient body fluid selected from urine, nasal secretions, nasal washes, bronchial lavages, bronchial washes, spinal fluid, sputum, gastric secretions, seminal fluid, other reproductive tract secretions, lymph fluid, whole blood, serum, or plasma. In addition, the disclosed workflow is integrated with clinical contexts such as transplant patients and anti-fungal infection risk assessment.
Claims Coverage
The independent claim covers a PCR-based DNA identification workflow for specifically identifying Aspergillus terreus in patient tissue or patient body fluid using a probe consisting of SEQ ID NO: 1. The dependent claims refine the inventive concept with specific primer/probe elements, real-time PCR, specified amplified DNA regions, enumerated specimen body fluids, and a clinical step involving anti-fungal agent administration.
Probe consisting of SEQ ID NO: 1 for species-specific hybridization
Hybridizing a probe to the extracted and amplified DNA to specifically identify the Aspergillus terreus fungal species, wherein the probe consists of SEQ ID NO: 1; and specifically identifying the Aspergillus terreus fungal species.
Forward and reverse primers for DNA amplification
Using a forward primer (SEQ ID NO: 2) and a reverse primer (SEQ ID NO: 3) to amplify DNA.
Real-time PCR amplification
Performing the method using real-time PCR.
Amplified DNA comprising internal transcribed spacer regions of nuclear ribosomal DNA
Having the amplified DNA consist of internal transcribed spacer regions of nuclear ribosomal DNA.
Enumerated patient body fluids for specimen selection
Using a patient body fluid selected from urine, nasal secretions, nasal washes, bronchial lavages, bronchial washes, spinal fluid, sputum, gastric secretions, seminal fluid, other reproductive tract secretions, lymph fluid, whole blood, serum, or plasma.
Administration of an anti-fungal agent
Further including administering an anti-fungal agent to the patient.
Overall, the claim set centers on extracting and recovering Aspergillus terreus DNA from patient tissue or specified body fluids, amplifying it, and specifically identifying the species by probe hybridization using SEQ ID NO: 1, with optional refinements including specific primers (SEQ ID NO: 2 and SEQ ID NO: 3), real-time PCR, a defined amplified target region (internal transcribed spacer regions of nuclear ribosomal DNA), enumerated specimen types, and adding anti-fungal agent administration.
Stated Advantages
Provides specific identification of Aspergillus terreus fungal species in patient tissue or patient body fluid using a probe consisting of SEQ ID NO: 1.
Supports use of probe formats including fluorescent labeling and bead dyed with a fluorochrome and supports real-time PCR.
Includes specimen flexibility by enumerating multiple patient body fluids suitable for the method.
Documented Applications
Identifying Aspergillus terreus in patient tissue or patient body fluid by extracting and recovering Aspergillus terreus DNA, amplifying it, hybridizing a probe (SEQ ID NO: 1), and specifically identifying the fungal species.
Integrated diagnostic context including transplant patients and anti-fungal infection risk.
Use of the method in workflows that can include administering an anti-fungal agent to the patient.
Interested in licensing this patent?