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A method of detecting microorganisms in blood

A technology of microorganisms and blood, applied in the biological field, can solve the problems of negative test results and low CO2 levels, and achieve the effects of great application value, high positive detection rate, and easy observation

Inactive Publication Date: 2016-03-02
SHANGHAI FOSUN LONG MARCH MEDICAL SCI CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But this bacteria-based CO 2 The detection method will be because some microorganisms (such as non-fermenting bacteria) that cannot utilize sugar metabolism do not produce CO 2 or produced CO 2 The amount is extremely low, resulting in a negative assay result

Method used

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  • A method of detecting microorganisms in blood
  • A method of detecting microorganisms in blood
  • A method of detecting microorganisms in blood

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1 (the present invention has used TTC as the indicator method of detecting microorganism)

[0025] (1) Take 25mL of polydimethylsiloxane, 750μL of ethyl n-laurate, 100μL of dibutyltin dilaurate, 1.25mL of cyclohexane and 0.5g of TTC (TTC was first dissolved in 0.5g of single distilled water) in Mix well in the container, add to the bottom of the blood culture bottle, solidify at 35°C, and prepare a silicone rubber oxygen-enriched membrane;

[0026] (2) Add 40mL broth medium and 4g732 cation exchange resin pellets (manufactured by Sinopharm Group) to the blood culture bottle, and sterilize at 121°C for 20min;

[0027] (3) Blood samples from patients with fever (≥38°C) or hypothermia (≤36°C), chills, and signs of leukocytosis were collected in the inpatient department of East China Hospital in Shanghai for experiments. Add 10 mL of blood sample to the blood culture bottle;

[0028] (4) Place the blood culture bottle with the blood sample in the BACTEC9050 aut...

Embodiment 2

[0030] Embodiment 2 (the present invention has used TTC as the indicator method of detecting microorganism)

[0031] (1) Take 35mL of broth medium, 2g of AmberliteXAD-4 non-ionic macroporous resin particles (produced by Rohm and Haas Company) and 1.05g of TTC (TTC is firstly dissolved in 1.05g of single distilled water) in a container and mix evenly, add Put it into a blood culture bottle and sterilize it at 121°C for 20 minutes;

[0032] (2) Fever (≥38°C) or hypothermia (≤36°C), chills, and leukocytosis (cell density greater than 10.0×10) were collected in the inpatient department of Shanghai Huadong Hospital. 9 Individual / L) blood samples from patients with signs were used for the experiment. Add 5 mL of blood sample to the blood culture bottle;

[0033] (3) Place the blood culture bottle with the blood sample in the BACTEC9050 automatic blood culture instrument of BD Company of the United States to incubate at 37°C, and monitor the culture medium at a distance of 2-3cm fr...

Embodiment 3

[0035] Embodiment 3 (colorimetric method)

[0036] (1) Take the "Zhuhai Deere aerobic blood culture bottle (colorimetric method)" for testing.

[0037] (2) Fever (≥38°C) or hypothermia (≤36°C), chills, and leukocytosis (count greater than 10.0×10) were collected in the inpatient department of Shanghai Huadong Hospital. 9 / L) blood samples from patients with signs were used for the experiment. Blood samples were collected by clinical microbiology laboratory technicians from clinical patients. Add 5 mL of blood sample to the blood culture bottle;

[0038](3) Place the blood culture bottle with the blood sample in the BACTEC9050 automatic blood culture instrument of BD Company of the United States to incubate at 35°C, and test it with the supporting detection equipment of the BACTEC9050 automatic blood culture instrument;

[0039] (4) After culturing for 62 hours, the detection signal shows that the light reflection intensity exceeds the threshold (such as image 3 ), and the...

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Abstract

A method of detecting microorganisms in blood is provided. The method includes (1) a step of directly adding 2,3,5-triphenyltetrazolium chloride (TTC) into a blood culture flask, or a step of mixing 2,3,5-triphenyltetrazolium chloride into a silicone rubber oxygen permeable membrane to obtain a solid indicator and fixing the solid indicator to the bottom of the blood culture flask, with the concentration of the TTC being 0.01-0.03 g / mL, (2) a step of adding a blood medium into the blood culture flask, and sterilizing at 121 DEG C for 20 min, (3) a step of adding a blood sample into the blood culture flask, and culturing at 35 DEG C for 0-5 days, and (4) a step of detecting intensity of reflected light of an index strip in the blood culture flask, mapping an intensity curve of the reflected light, and determining a microorganism growth result according to the curve. The method is advantaged by low costs, simple operation processes, short detection time, high positive detection rates, and the like in clinical application, and is high in application value.

Description

technical field [0001] The invention relates to biotechnology, in particular to a method for detecting microorganisms, in particular to a method for detecting microorganisms in blood. Background technique [0002] Blood culture is one of the important detection items in clinical laboratories. Since the mortality rate of bacteremia and sepsis is as high as 30-50%, accurate and timely culture results are of great significance for clinical detection and treatment. At present, the positive detection rate of bacteria in blood culture in China is only 10%. The possible reasons are: 1. Low bacterial content in blood; 2. Antibacterial substances in blood; 3. Application of antibiotics; 4. Culture conditions limits. Blood culture testing products currently on the market mainly use CO 2 The main detection methods are radioactive labeling technology, fluorescence detection technology and spectrophotometric technology. Bacteria produce CO during their growth and metabolism 2 , the p...

Claims

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Application Information

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IPC IPC(8): G01N21/31G01N1/28
Inventor 王龙胡维冉苇
Owner SHANGHAI FOSUN LONG MARCH MEDICAL SCI CO LTD
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