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Separation reagent, preparation method and applications of separation reagent, method for separating bacteria and gel extraction tube

A reagent and gel technology, which can be used in bacteria, separation of microorganisms, etc., can solve the problems of long time, low yield, and complicated operation.

Active Publication Date: 2021-05-11
珠海恒屹生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this method is cumbersome and time-consuming to operate, and more bacterial cells are lost in differential centrifugation, so the harvest rate is low, so few people currently use it.
[0007] Secondly, the membrane filtration method was used in the early research, but it was quickly abandoned because the medium composition of the activated carbon blood culture bottle is complex, including not only activated carbon with different particle sizes, but also broken blood cell fragments, as well as protein colloids, which are easy to Blocking the filter membrane and not being able to remove activated carbon well also leads to inaccurate and time-consuming bacterial detection

Method used

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  • Separation reagent, preparation method and applications of separation reagent, method for separating bacteria and gel extraction tube
  • Separation reagent, preparation method and applications of separation reagent, method for separating bacteria and gel extraction tube
  • Separation reagent, preparation method and applications of separation reagent, method for separating bacteria and gel extraction tube

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Experimental program
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preparation example Construction

[0104] The embodiment of the present invention also provides a preparation method of the above separation reagent, comprising:

[0105] The gel that the embodiment of the present invention adopts is self-made gel, and the synthesis of gel comprises: Dimethyl silicone oil, hydroxyl silicone oil, silicon dioxide and silane coupling agent are (11-17) according to mass ratio: (0.8-3 ): (2-5): (0.2-1) after mixing in a ratio of cross-linking reaction to form a silicone polymer;

[0106] Preferably, the operation steps of the crosslinking reaction are as follows: after mixing dimethyl silicone oil and hydroxy silicone oil and heating to 100-120°C, adding silicon dioxide and silane coupling agent while stirring, and then stirring for 8-12 hours, the reaction ends Then vacuum defoaming at 150-170°C for 3-5 hours. Adopting the above method can ensure the density of the prepared gel and ensure the separation effect of the gel.

[0107] It should be noted that the gel can also be purch...

Embodiment 1- Embodiment 3

[0121] In terms of parts by weight for preparing 100 extraction tubes, each tube is 1ml.

[0122] The proportioning of the second component in the separation reagent of embodiment 1-embodiment 3 is as follows:

[0123]

[0124] It should be noted that there is no additional value in the table, and the unit is g.

[0125] The first component in the separation reagent of embodiment 1-embodiment 3 is organosilicon polymer, and the proportioning of first component and second component in the separation reagent of embodiment 1-embodiment 3 is as follows:

[0126]

[0127] The raw material ratio of the organosilicon polymer of the first component in the separation reagent of embodiment 1-embodiment 3 is as follows:

[0128]

[0129] The preparation method of the separating reagent of embodiment 1-embodiment 6 is identical, comprises:

[0130] Preparation method of the second component:

[0131] Hemolyzing reagents, buffers, and centrifugation gradients are mixed and ster...

Embodiment 4

[0137] Example 4: The first component is 10 grams of a gel with a trade name of “Anti-Radiation Organosilicon Serum Separation Gel” from Wuhan Desheng Biochemical Technology Co., Ltd., with a density of 1.042.

[0138] The second component includes 2 mg phospholipase C (Clostridium perfringens source phospholipase C, CAS: 9001-86-9), 0.01 × 10 -3 Gram Tween 80 and 0.6% PIPES buffer, ion regulator are the same as in Example 1, the total amount of the second component is 100 milliliters, and its pH is 7.2. 0.1 gram of the first component was added for every milliliter of the second component.

[0139] The preparation method of the separation reagent of embodiment 4 is basically the same as the preparation method of the separation reagent of embodiment 1.

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Abstract

The invention discloses a separation reagent, a preparation method and applications of the separation reagent, a method for separating bacteria and a gel extraction tube, and relates to the technical field of biological detection. The separation reagent includes a first component and a second component; the first component includes, by weight in parts, 10-30 parts of gel; and the second component includes 2*10<-3>-2.0 parts of a hemolytic agent, (0.01-0.5)*10<-3> parts of a centrifugal gradient regulating agent and a buffering agent, and the pH of the second component can be 7.2-7.6 through the using amount of the buffering agent. The separation reagent can well treat the nutrient solutions cultured by enriched blood to separate substance such as culture media, blood cells and activated charcoal in culture cultured by bacteria and blood, so that pure bacteria with high concentration can be obtained through quick separation, bacterial detection can be directly carried out after enriched blood culturing, therefore, detection speed can be accelerated, and the influences of the substances such as the culture media, the blood cells and the activated charcoal on bacterial detection can be reduced.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a separation reagent, a preparation method, an application thereof, a method for separating bacteria and a gel extraction tube. Background technique [0002] Blood culture is an artificial culture method to identify pathogenic bacteria. For the etiological diagnosis of bacteremia, fungemia, sepsis and sepsis. In particular, infections in meningitis, sepsis, thoracic and abdominal cavities and other sterile sites are critical infections. The traditional diagnosis method is: blood culture is enriched and then subcultured on solid medium, and after 1-2 days of subculture, directional identification is carried out , followed by system identification and drug susceptibility testing, it often takes 3-4 days to complete the testing and issue a report. However, more than 99% of the positive blood cultures are single bacteria, and the separation and purification effect of s...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/02
CPCC12N1/20C12N1/02
Inventor 卢先雷
Owner 珠海恒屹生物科技有限公司
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