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Method for detecting microorganisms in fuel oil by using bioluminescence method and special detection rod thereof

A detection rod and fluorescence method technology, applied in the field of biochemical analysis and detection, can solve the problems of low accuracy, poor identification, high cost, etc., and achieve the effects of high stability, improved detection speed, and consistent reaction time

Active Publication Date: 2021-01-22
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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  • Method for detecting microorganisms in fuel oil by using bioluminescence method and special detection rod thereof
  • Method for detecting microorganisms in fuel oil by using bioluminescence method and special detection rod thereof
  • Method for detecting microorganisms in fuel oil by using bioluminescence method and special detection rod thereof

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Experimental program
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Effect test

Embodiment 1

[0070] The embodiment of the present application discloses a detection stick swab handle which is convenient for fastening and releasing the swab, including: a swab main pipe 1, the surface of the swab main pipe 1 is provided with a guide thread 2, and the surface of the swab main pipe 1 is movably installed with a swab The sub-handle 10, the bottom of the swab main pipe 1 is fixedly installed with a supporting plate 3, the bottom of the supporting plate 3 is fixedly installed with an auxiliary pipe 4, the swab main pipe 1 and the auxiliary pipe 4 are connected, and the bottom of the auxiliary pipe 4 is fixedly installed with an installation sleeve 5 , the bottom of the mounting sleeve 5 is screwed with a detection tube 6, and the outer side of the detection tube 6 is movably socketed with a microtube cover 15, and the microtube cover plays a light-proof effect. A first mixing chamber and a second mixing chamber are respectively arranged up and down in the detection tube 6, and...

Embodiment 2

[0073] combine figure 1 and Figure 5 , the positioning part 9 includes a sliding block 91 sliding in the inner cavity of the long sliding hole 12 and a chute 94 provided on the inner wall of the guide frame 7, the two sides of the sliding block 91 are welded with ball sleeves 92, and the inner cavity of the ball sleeve 92 is movable. Balls 93 are installed, and one side of the sliding block 91 is threaded with a helical positioning pin 95 that matches the positioning groove 8. Through the setting of the positioning piece 9, when the position of the swab handle 10 moving down needs to stop at a suitable position , the user can slide the sliding block 91 up and down, the sliding block 91 drives the ball sleeve 92 to slide in the inner cavity of the chute 94, and at the same time, the ball 93 will roll against the inner wall of the chute 94 and move to a suitable position corresponding to the positioning groove 8 , then tighten the twist block 96, and insert the helical positio...

Embodiment 3

[0075] Preparation of liquid reagents in test tubes:

[0076] 1) Weigh 4.48g of trimethyl amino acid, 0.6g of copper sulfate, 0.146g of EDTA, 100mg of bovine serum albumin, and 77mg of dithiothreitol, and add them into 600ml of water.

[0077] 2) Add 4ml of 10U / ml apyrase and 1ml of 10U / ml adenosine phosphate deaminase, adjust the pH to 7.8 with 10% sodium hydroxide, and distill the volume to 1 liter with distilled water. Make the activity respectively 0.04U / ml and 0.01U / ml.

[0078] 3) Aliquot it into each assay, 1ml per tube.

[0079] Preparation of powder reagents in test tubes:

[0080] 1) Weigh 1 μg of luciferase, 7.254ng of sodium D-luciferin, 1.3146 μg of magnesium sulfate and 0.25mg of octadecyltrimethylammonium chloride, and mix well.

[0081] Preparation of detection tubes:

[0082] 1) Add the powdered reagent into a 2ml screw-top plastic detection tube, and seal 1ml of the liquid reagent in the aluminum foil on the upper part of the detection tube.

[0083] Assem...

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Abstract

The invention provides a method for detecting microorganisms in fuel oil by using a bioluminescence method and a special detection rod thereof. According to the detection method disclosed by the invention, a mixture of adenosine triphosphate diphosphatase, adenosine phosphate deaminase and copper sulfate is used as a free ATP removing agent so that free ATP in a sample can be effectively removed;octadecyl trimethyl ammonium chloride is adopted as a cell lysis agent, a lysis speed is high, the lysis is complete, and the remaining ATP hydrolase can be irreversibly inactivated; luciferase is a freeze-dried powder and can be stored at a normal temperature so that the storage cost is saved; the rod part of the special matched detection rod adopts a thread design so that the stability and consistency of a result are ensured; and fluorescent aggregation reaction balls are contained in a detection tube and used for aggregating fluorescent signals, and the detection sensitivity, stability andaccuracy are improved.

Description

technical field [0001] The invention relates to a preparation method and a measurement method for a detection rod used for the rapid detection of microorganisms in a fuel sample by an ATP biofluorescence method, belonging to the field of biochemical analysis and detection. Background technique [0002] According to statistics, 50% of engine failures are caused by poor fuel cleanliness. Microbes are an important factor affecting the cleanliness of fuel, and have a significant impact on fuel storage and operation safety. Since 1930, people have gradually realized a series of hazards caused by the growth and reproduction of microorganisms in fuel, and the problem of microbial contamination of fuel has begun to attract attention. Microbial pollution is a major threat to the aviation industry, marine industry and transportation industry. A large number of microorganisms and microbial secretions condense into sticky lumps or flocs that can block oil filters, oil pumps, fuel regul...

Claims

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Application Information

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IPC IPC(8): G01N21/64
CPCG01N21/6428
Inventor 刘龙飞
Owner BEIJING ZHONGJIAN BAOTAI BIOTECH
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