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Methods and compositions for detecting microorganisms

A technology of microorganisms and compositions, applied in biological testing, biological material analysis, measuring devices, etc.

Inactive Publication Date: 2011-02-23
3M INNOVATIVE PROPERTIES CO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Although various methods are currently available for the detection and enumeration of fungal microorganisms, there is a need for faster and more sensitive methods for the detection and enumeration of fungal microorganisms

Method used

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  • Methods and compositions for detecting microorganisms
  • Methods and compositions for detecting microorganisms
  • Methods and compositions for detecting microorganisms

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0100] Rabbit anti-zymosan antibody recognizes zymosan by indirect ELISA .

[0101] A. General Materials and Methods

[0102] By adding 3.03g Na 2 CO 3 and 6.0 g NaHCO 3 Mix in 1000 mL of distilled water to prepare coating buffer (0.1 M bicarbonate / carbonate buffer, pH 9.6) as a 1X stock solution. Phosphate buffered saline (PBS, 155 mM NaCl in 10 mM phosphate buffer, pH 7.4) was prepared from 1OX stock solutions from Biosource (Rockville, MD, USA). BD Falcon Microtest TM 96-well enhanced surface ELISA plates were obtained from BD Biosciences (Bedford, MA, USA). All procedures were performed at room temperature unless otherwise stated. All ELISA wash programs consisted of three consecutive wash volumes of 200 μl / well, and all washes were performed with PBS buffer. The horseradish peroxidase chromogenic substrate 3,3',5,5'-tetramethylbenzidine (TMB) was obtained from Pierce Biotechnology (Rockford, IL, USA). 2M sulfuric acid was prepared from concentrated mother liq...

example 2

[0114] Biotinylated anti-zymosan antibody via biotin-streptavidin ELISA recognizes yeast poly sugar .

[0115] A. General Materials and Methods

[0116] Coating buffer, PBS, ELISA plates, incubation and washing procedures were performed as described in Example 1. Alkaline phosphatase, chromogenic substrate, disodium p-nitrophenyl phosphate (pNPP, catalog #37621 ) were obtained from Pierce Biotechnology. 2N sodium hydroxide was prepared from a stock concentration of 5M and obtained from Sigma-Aldrich. Bovine serum albumin (BSA) was used in this experiment as a 1% solution in PBS and was obtained from Sigma-Aldrich. Alkaline phosphatase (AP)-conjugated Immunopure streptavidin was obtained from Pierce Biotechnology.

[0117] Zymosan antigen was prepared and stored as described in Example 1.

[0118] Antibodies used in this experiment included rabbit anti-Zymosan (cat. no. Z2850) from Molecular Probes. Rabbit anti-Zymosan was biotinylated using the EZ-Link Sulfo-NHS B...

example 3

[0125] Liposomes recognize zymosan through anti-zymosan antibody interactions .

[0126] A. General Materials and Methods

[0127] Coating buffer, PBS, ELISA plates, incubation and washing procedures were performed as described in Example 1. Bovine serum albumin (BSA) was used in this experiment as a 1% solution in PBS and was obtained from Sigma-Aldrich. ImmunoPure Streptavidin was obtained from Pierce Biotechnology.

[0128] Liposomes were lysed with 0.25% TRITON X-100 aqueous solution. The horseradish peroxidase chromogenic substrate 3,3',5,5'-tetramethylbenzidine (TMB) was obtained from Pierce Biotechnology. 2M sulfuric acid was prepared as described in Example 1 from the concentrated mother liquor.

[0129] Zymosan antigen was prepared and stored as described in Example 1.

[0130] A biotinylated rabbit anti-zymosan antibody was prepared as described in Example 2.

[0131] B. HRP liposome synthesis

[0132] Measure 1.25 mL of 20 mg / mL 1,2-dipalmitoyl-sn-glyce...

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Abstract

Methods of analyzing a sample for target microorganisms of interest are described. In particular, the methods are useful for detecting a variety of yeast and mold microorganisms based on the presence of a ubiquitous cell wall component, such as zymosan. Methods of analyzing a sample for fungal microorganisms using liposomes and / or culture media are also described.

Description

[0001] Cross references to related patent applications [0002] This application claims the benefit of US Provisional Application Serial No. 61 / 028,898, filed February 14, 2008, which is incorporated herein by reference. Background technique [0003] Fungal microorganisms such as yeasts and molds are ubiquitous in nature. Although fungal microorganisms such as yeasts and molds are useful in areas such as the production of antibiotics or enzymes, they are known to cause food spoilage, certain species are also known to produce toxins or cause infections in humans or animals. [0004] Fungal microorganisms comprise a very large and diverse group of chemoheterotrophic microorganisms. There are about 1500 species of yeast. Most yeasts exist as single-celled organisms that reproduce by budding. Some species exist in a multicellular form, while others reproduce by binary fission. Molds include filamentous fungi that form long, multicellular hyphae. Molds reproduce by forming sm...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53
CPCG01N33/5308G01N33/569G01N2333/39G01N2469/10G01N2400/24
Inventor 史蒂芬·B·罗斯科斯特凡妮·J·莫勒曼基里·T·克希尔萨加尔
Owner 3M INNOVATIVE PROPERTIES CO
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