Magnetic resonance spectroscopy to identify and classify microorganisms

A microbial and magnetic resonance technology, applied in the field of magnetic resonance spectroscopy analysis for microbial identification and classification, can solve problems such as time-consuming, expensive, and a lot of work

Inactive Publication Date: 2005-02-16
NAT RES COUNCIL OF CANADA +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional methods are usually time-consuming
When some physiological experiments are required to make a definitive identification, for some species of yeast such as Candida, in which only few phenotypic and chemotypic differences exist, these experiments may be difficult or even unsuccessful for identification
Genetics methods are more accurate but work-intensive and expensive

Method used

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  • Magnetic resonance spectroscopy to identify and classify microorganisms
  • Magnetic resonance spectroscopy to identify and classify microorganisms
  • Magnetic resonance spectroscopy to identify and classify microorganisms

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Embodiment 1: Bacterial detection

[0046] 1. Bacteria Storage and Culture

[0047] Isolates were from deposits of the Center for Infectious Diseases and Microbiology (CIDM) and the American Type Culture Collection, Institute of Clinical Pathology and Medical Sciences, Sydney, or the most recent clinical isolate from the Clinical Qualification Laboratory, CIDM Laboratory Services . Stored bacteria were suspended in 10% glycerol nutrient broth at -70°C. Horse blood agar (HBA) was prepared by adding sterile horse blood to autoclaved blood agar medium (Oxoid (UK) or Amyl Media (Australia)). The bacteria reduced from the stored bacteria were subcultured on 5% horse blood agar and incubated at 37°C, 5% CO 2 Conditioned for 18-24 hours. Newly grown bacteria and bacteria subcultured on HBA after storage were inoculated on duplicate HBA dishes, and incubated at 37°C for 18-24 hours, and then incubated at room temperature (20-30°C) before spectroscopic detection. ) stored f...

Embodiment 2

[0092] Example 2: Using MRS to Discriminate Cryptococcal and Glioma in Mouse and Cell Culture

[0093] 1. Preamble

[0094] Characterization of clinical isolates of Cryptococcus neoformans and glioma cell lines in culture and in experimental mice using MRS. 1D and 2D collections from in vitro cultured fungi (16 strains of Cryptococcus neoformans, 3 species of Candida albicans, 3 species of Aspergillus fumigatus, 3 species of Saccharomyces cerevisiae) and C6 glioma cell lines 1 H MR spectroscopy. Brain biopsies were taken from healthy mice and experimentally infected mice or gliomas (19 healthy brains, 19 cryptococci and 20 gliomas) and were analyzed using homogeneous and heterogeneous nuclei 2D correlation spectroscopy (COSY, TOCS, 1 H, 13 C-HSQC and HMBC) for unambiguous signal assignment between cell suspensions and tissue samples. The results indicated that the MR spectra of C. neoformans and C. neoformans were dominated by the resonance of the disaccharide a, a-trehalose...

Embodiment 3

[0133] Example 3: Identification of pathogenic fungi

[0134] 1. Microorganisms

[0135] 205 cultures of pathogenic yeasts Candida albicans, Candida parapsilosis, Candida tropicalis, Candida krusei, and Candida glabrata were grown on Sabouraud dextrose agar at 30°C for 48 hours. 69 and 70 isolates of the pathogenic yeast Cryptococcus neoformans var. neoformans and var. gattii, respectively, were cultured on Sabouraud dextrose agar at 37°C for 48 hours. Yeasts were biochemically identified using the API 20C AUX system (BioMerieux, Marcy 1'Etoile, France). Cryptococci were biotyped and serotyped (Crypto Check agglutinationtest, Iatron Labs). In addition, fingerprint PCR was used to compare the serotypes of the corresponding species. Colonies were scraped from Petri dishes and suspended in PBS / D immediately prior to MR experiments 2 O middle.

[0136] 2. MR spectrum detection

[0137] with assembly with 5mm { 1 H, 13 C) MR spectra were acquired with a Bruker Avance 360MHz ...

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Abstract

A statistical classifier identifies microorganisms, such as bacteria and fungi, using magnetic resonance spectroscopy, with multivariate analysis. The bacteria may include species within Staphylococcus, Enterococcus and Streptococcus. The fungi may include pathogenic yeasts including species with Candida and Cryptococcus.

Description

[0001] Refer to related application [0002] This application claims priority to US Provisional Application No. 60 / 270,367, filed February 21, 2001. Background technique [0003] The present invention relates to the identification and classification of microorganisms such as bacteria and fungi using magnetic resonance spectroscopy and multivariate analysis. [0004] Throughout the application, individual references are placed in parentheses. The entirety of these documents are hereby incorporated by reference into this application in order to more fully describe the state of the art to which this invention pertains. A bibliography of these documents can be found at the end of the application, just before the claims. [0005] Microtaxonomic classification of microorganisms involves grouping microorganisms into groups based on characteristics such as detection of various metabolites / compounds of the microbial cells or analysis of genetic material. "Gene trees" derived from se...

Claims

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Application Information

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IPC IPC(8): G01R33/465C12M1/34C12M1/42C12Q1/06C12Q1/10C12Q1/14G01R33/32G01R33/46
CPCG01R33/465G01R33/4625
Inventor 雷蒙德·L·召莫尔尧伊塔尼亚·佐雷尔卡罗琳·E·芒福德乌韦·希默尔赖希罗杰·伯恩
Owner NAT RES COUNCIL OF CANADA
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