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Reagent and method for separating bacteria from positive blood and method for identifying bacteria

A positive blood and reagent technology, applied in the field of bacterial separation and identification in blood, can solve the problems that the accuracy of bacterial identification needs to be further improved, and achieve the effects of shortening the time spent, high identification accuracy, and improving work efficiency

Inactive Publication Date: 2018-01-26
PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, in the above prior art, the accuracy of bacterial identification in blood needs to be further improved

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] The reagent for separating bacteria from positive blood consists of: saponin solution (agent A), cleaning solution (agent B), formic acid and acetonitrile (agent C).

[0050] Among them, the concentration of saponin solution is 5% (add 5 grams of saponin in 100mL water and mix well), and the consumption is 40 microliters; the cleaning solution is deionized water, and the consumption is 1000 microliters; the concentration of formic acid is 100%, and the concentration of acetonitrile 99.8%, the dosage is 20 microliters each. Volumes are based on 200 microliter samples.

[0051] Each component in the above reagents can also be stored in a plurality of separate storage spaces, and can be processed into a kit, and the kit is divided into multiple storage spaces to store the components. In this embodiment, the reagents are made into kits. There are 2000 microliters of agent B in the kit, and these 2000 microliters of agent B can be stored in two spaces or in one storage spa...

Embodiment 21

[0089] The reagent for isolating bacteria from positive blood is different from Example 1 in that the reagent A for lysing blood cells is a mixture of SDS and saponin solution.

[0090] In this example, Escherichia coli, Staphylococcus aureus, Enterococcus faecalis, Klebsiella pneumoniae, Acinetobacter baumannii or Staphylococcus epidermidis were added to the sterile blood, identified after cultivation, and repeated 10 times In the experiment, for each type of bacteria, the identification accuracy rate is above 95% on average.

[0091] Acinetobacter baumannii, Staphylococcus epidermidis and Klebsiella pneumoniae were added to the sterile blood, and identified after culture. Repeated experiments were repeated 10 times, and the average identification accuracy rate was over 95%.

[0092] Enterococcus faecalis, Acinetobacter baumannii, Staphylococcus epidermidis and Klebsiella pneumoniae were added to the sterile blood, and identified after culture. The experiment was repeated 10 ...

Embodiment 22

[0095] The reagent for isolating bacteria from positive blood is different from Example 1 in that the A agent for lysing blood cells is SDS and saponin solution. When operating, first use one of them to lyse blood cells, and then use the other to lyse blood cells again.

[0096] In this example, Escherichia coli, Staphylococcus aureus, Enterococcus faecalis, Klebsiella pneumoniae, Acinetobacter baumannii or Staphylococcus epidermidis were added to the sterile blood, identified after cultivation, and repeated 10 times In the experiment, for each type of bacteria, the identification accuracy rate is above 95% on average.

[0097] Acinetobacter baumannii, Staphylococcus epidermidis and Klebsiella pneumoniae were added to the sterile blood, and identified after culture. Repeated experiments were repeated 10 times, and the average identification accuracy rate was over 95%.

[0098] Enterococcus faecalis, Acinetobacter baumannii, Staphylococcus epidermidis and Klebsiella pneumoniae...

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PUM

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Abstract

The invention provides a reagent for separating bacteria from positive blood. The reagent comprises an agent A, an agent B and an agent C, wherein the agent A is used for dissolving the hemocyte froma blood sample of a blood culture; the agent B is used as a cleaning agent; the agent C is used for breaking the pathogenic bacteria cytoderm and promoting the protein release in the cells; and the agent A is at least one of saponin solution and SDS or other surface active agent capable of cracking the hemocyte. The invention also provides a method for separating bacteria from positive blood and the method for identifying bacteria. The method comprises the following steps: cracking the hemocyte by using the agent A; cleaning the cell fragments and impurities by using the agent B; breaking thepathogenic bacteria cytoderm by using the agent C, releasing the bacteria protein, taking the supernate containing the bacteria protein and settling a sample to be identified; dropping the sample to be identified into a vacant side on a target plate; drying under room temperature and then covering a substrate on the sample; drying under room temperature; and putting onto a machine for detecting. The bacteria can be quickly identified, the result can be acquired within half an hour, the detection result is high in reliability and the interference is less.

Description

technical field [0001] The invention belongs to the field of bacteria isolation and identification in blood. Specifically, the invention provides a reagent and method for isolating bacteria from positive blood and a bacteria identification method. Background technique [0002] Septicemia refers to the acute systemic infection caused by pathogenic bacteria or opportunistic pathogenic bacteria invading the blood circulation, growing and multiplying in the blood, and producing toxins. For the diagnosis and treatment of sepsis, the blood of the patient needs to be cultured for bacteria, and then the bacteria can be identified before the patient can be given medication according to the identification result. [0003] Common pathogenic bacteria include Staphylococcus aureus, Escherichia coli, Streptococcus pneumoniae or Klebsiella pneumoniae, etc. The pathogenic bacteria of children and immunocompromised persons can be Staphylococcus epidermidis. [0004] With the existing reagen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/02C12N1/20G01N27/62
Inventor 肖盟冯立平徐英春范欣杨启文王瑶王贺孙宏莉
Owner PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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