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Assay for the identification of microorganisms

A microbial and electrochemical technology, applied in the field of analysis for identifying microorganisms, which can solve problems such as unreliability and time-consuming

Inactive Publication Date: 2007-02-14
RAPID LAB MICROSYST INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Further, existing tests are time consuming and often unreliable

Method used

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  • Assay for the identification of microorganisms
  • Assay for the identification of microorganisms
  • Assay for the identification of microorganisms

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Respiratory cycle activity in the presence of effector compounds

[0029] In this example, the assay uses the following steps: (1) The method involves mixing 200 μL of bacterial culture with 1300 μL of buffer (with or without effector and 5 μM DCIP) at a fixed temperature (35° C.) The electrode was mixed in the electrochemical cell for a fixed time (10 min); (2) followed by the addition of mediator (500 μL of 0.2M potassium ferricyanide) at a fixed temperature (35°C) for a fixed time (10 min); and (3) The current is measured at a fixed voltage difference (-100mV) and for a fixed time (120sec). Ferricyanide solution was added such that the final concentration of ferricyanide was 50 mM.

[0030] In this configuration, the analysis uses approximately 0.017cm each 2 surface area of ​​the two platinum electrodes. The measured current is proportional to the concentration of the least concentrated redox pair species in solution. In the present system, there is a large exce...

Embodiment 2

[0042] Distinguish between Gram-positive and Gram-negative bacteria

[0043] The following steps were used for each experiment: (1) Preheat 150 μL aliquots of buffer containing 1 mM glucose (glc) with or without 5 μM DCIP at a fixed temperature (35° C.) for a fixed time (4 min); (2 ) then add 50 μL of bacterial suspension to the sample at a fixed temperature (35°C) for a fixed time (10 min); (3) then add mediator (50 μL 0.4M ferricyanide Potassium) for a fixed time (10 min); and (4) current was measured at a fixed temperature (35° C.) and for a fixed time (120 sec) at a fixed voltage difference (100 mV) between two platinum electrodes. The final ferricyanide concentration in the sample prior to reaction with the microorganism was 40 mM.

[0044] In this configuration, the analysis uses approximately 0.03cm each 2 surface area of ​​the two platinum electrodes. The measured current depends on the concentration of the least concentrated redox species in solution. In our syste...

Embodiment 3

[0051] Discrimination of 10 microbial strains using pattern recognition by principal component analysis of respiratory activity data

[0052] In this example, individual microorganisms were subjected to 22 different effector compounds in addition to control measurements. The following steps were used for each experiment: (1) Preheat 150 μL aliquots of buffer containing the effector compound and 5 μM DCIP at a fixed temperature (35° C.) for a fixed time (4 min); (2) subsequently at a fixed temperature ( Add 50 μL of bacterial suspension to the sample at 35°C) for a fixed time (10 min); (3) then add mediator (50 μL 0.4M potassium ferricyanide) at a fixed temperature (35°C) for a fixed time (10 min); and (4) current was measured at a fixed temperature (35° C.) and for a fixed time (120 sec) at a fixed voltage difference (100 mV) between two platinum electrodes. The final ferricyanide concentration in the sample prior to reaction with the microorganism was 40 mM. For each microo...

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Abstract

A method for the phenotypic identification of microorganisms is provided. The method is based on the evaluation of the effects of various compounds (effectors) on the respiratory cycle activity of microorganisms. Measurements are based upon the ability of the microorganism to transport electrons to an external chemical oxidant (a mediator) that is added to the microorganism sample. The mediator interacts with the terminal components of the respiratory pathway and the extent of its consumption is related to the ability of the microorganism to respire. The consumed mediator is subsequently measured electrochemically. Electrochemical signals which are generated in the presence or absence of an effector can be used to generate a signal pattern that is unique to an organism and can be used for identification.

Description

field of invention [0001] The present invention relates to assays for the identification of microorganisms. More particularly, the invention relates to the detection of prokaryotic and eukaryotic microorganisms in a multi-assay format for the rapid identification and characterization of cultures important for clinical, agricultural and environmental testing. Background of the invention [0002] Many species of microorganisms are harmful to humans or animals, so the correct identification of disease-causing pathogens is of great importance. Microbial diseases are a leading cause of death in many developing countries around the world. An increasing number of microbial pathogens have been identified as important food- and water-borne pathogens. [0003] Traditional microbial identification methods include a pre-enrichment step and a selective enrichment step, followed by morphological examination, biochemical screening, growth characterization, serotyping, and serological con...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/04G01N27/26C12N1/20
CPCC12Q1/04
Inventor 苏珊·R·米克凯尔森彼得·厄特尔道格·斯帕克斯利亚姆·奥黑根托马斯·曼保罗·乌尔里克
Owner RAPID LAB MICROSYST INC
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