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A method for detecting microorganisms in fuel oil by bioluminescent method and its special detection stick

A detection rod and fluorescence method, applied in the field of biochemical analysis and detection, can solve the problems of low detection sensitivity, inability to directly detect microbial cell ATP, and poor discrimination.

Active Publication Date: 2021-09-14
BEIJING ZHONGJIAN BAOTAI BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Existing fuel microbial detection pens using the ATP method have complex sample processing and cumbersome operation steps, and cannot directly detect ATP in microbial cells. The total ATP content and free ATP content need to be measured separately. The difference between the two is the The ATP of ATP; At the same time, the identification is poor; In addition, the cost of the existing fuel microbial detection pen is relatively high
[0005] Based on the above situation, it is urgent to develop a detection method based on bioluminescent reaction, which can efficiently remove the free ATP in the detection sample, and can effectively release the ATP in the microbial cells, and the cracking speed is fast. Due to the shortcomings of low detection sensitivity and low accuracy of the detection stick, a fuel microbial detection stick specially suitable for bioluminescent reactions has been developed to make its fluorescence reactions concentrated and stable.

Method used

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  • A method for detecting microorganisms in fuel oil by bioluminescent method and its special detection stick
  • A method for detecting microorganisms in fuel oil by bioluminescent method and its special detection stick
  • A method for detecting microorganisms in fuel oil by bioluminescent method and its special detection stick

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Effect test

Embodiment 1

[0070] In the present application, the present application discloses a detecting a baffle handle that facilitates the fastening of the swab, including: swab lead, the surface of the swab lead is opened, and the surface activity of the swab lead is installed. The sub-handle 10, the bottom of the swab main pipe 1 is fixedly mounted, and the bottom of the pallet 3 is fixedly mounted, and the swab main pipe 1 and the deputy tube 4 are connected, and the bottom portion of the sub-pipe 4 is fixedly mounted. 5 The bottom thread of the mounting sleeve 5 is mounted with a detection tube 6, the detection tube 6, the outer movable sleeve cover 15, the microtube cover acts as a light of light. The first mixing chamber and the second mixing chamber are respectively disposed in the detection tube 6, respectively, respectively, a liquid reagent and a powder reagent; the bottom of the detecting tube 6 is provided with a fluorescent aggregation reaction ball 61, and the fluorescent aggregation rea...

Embodiment 2

[0073] Combine figure 1 with Figure 5 The positioning member 9 includes a sliding block 91 sliding on the inner cavity of the long slide hole 12 and a sliding groove 94 on the inner wall of the guide frame 7, and both sides of the sliding block 91 are welded with a ball sleeve 92, and the rolling bead sleeve 92 The ball 93 is mounted, one side thread of the slide block 91 is connected to the screw positioning pin 95 that is fitted with the positioning groove 8, by the positioning member 9, when the position of the swab handle 10 is stopped at a suitable position The user can slide the slide block 91 up and down, the sliding block 91 drives the ball sleeve 92 to slide on the inner cavity of the chute 94, and the ball 93 can be scarled to the inner wall of the chute 94, move to the position of the corresponding positioning groove 8. When the twigent block 96 is tightened, the spiral position pin 95 is inserted into the positioning groove 8, so that the swab 10 can be positioned in a...

Embodiment 3

[0075] Preparation of liquid reagents in the tube:

[0076] 1) Weighing 4.48 g of trimethyl amino acid, 0.6 g of copper sulfate, 0.146 g of EDTA, 100 mg of bovine serum albumin, 77 mg of dithioseol, adding 600 ml of water.

[0077] 2) Add 4 ml of 10u / ml of triphosphate bispipanoase and 1 ml of 10u / ml of adenosyl phosphatase, adjusted pH to 7.8 with 10% sodium hydroxide, and set to 1 liters with distilled water. It is 0.04 U / mL and 0.01 U / mL, respectively.

[0078] 3) Dispen to each test, 1 ml per tube.

[0079] Detection reagent powder pipes prepared:

[0080] 1) Weigh 1μg luciferase, 7.254ng D- fluorescein sodium, 1.3146μg 0.25mg of magnesium sulfate and stearyl trimethyl ammonium chloride, and mix.

[0081] Preparation detection tube:

[0082] 1) The powder reagent was added 2ml plastic screw cap test tube, 1ml of liquid reagent sealed in aluminum foil in the upper portion of the test tube.

[0083] Detection rod assembly:

[0084] 1) A test tube, the swab shank and a thre...

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Abstract

The invention provides a method for detecting microorganisms in fuel oil by a bioluminescence method and a special detection stick thereof. The detection method of the present invention adopts the mixture of apyrase, adenosine phosphate deaminase and copper sulfate as the free ATP remover, which can effectively remove the free ATP in the sample; adopts octadecyltrimethylammonium chloride as the cell The cracking agent has a fast and complete cracking speed and can irreversibly inactivate the remaining ATP hydrolase; the luciferase is freeze-dried powder, which can be stored at room temperature to save storage costs; the special matching detection rod of the present invention adopts threaded Designed to ensure the stability and consistency of the results; the detection tube contains fluorescent aggregation reaction balls, which are used to gather fluorescent signals to improve the sensitivity, stability and accuracy of detection.

Description

Technical field [0001] The present invention relates to a method for preparing rapid detection and determination of ATP bioluminescence assay for detecting microorganisms in a sample of fuel rods for biochemistry analysis and detection belongs. Background technique [0002] According to statistics, 50% of engine failures are caused by poor cleanliness of the fuel caused. Microbes is an important factor affecting the cleanliness of the fuel, have a significant impact on the storage and safe operation of fuel. Since 1930, people have realized that fuel growth and reproduction of microorganisms in a series of hazards caused by microbial contamination of fuel began to receive attention. Microbial contamination is a major threat to the aviation industry, transport industry and marine industry, a lot of microorganisms, secretions condensed to a viscous slug or floes can be closed oil filter, oil pump, fuel control unit and other accessories, the engine direct impact normal oil, affect ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64
CPCG01N21/6428
Inventor 刘龙飞
Owner BEIJING ZHONGJIAN BAOTAI BIOTECH
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