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Bacterium identification reagent kit as well as preparation method and uses thereof

A kit and bacteria technology, applied in the field of bacteria identification kits and its preparation using the only carbon source for identification of bacteria, can solve the problems of low comparability of results, high price, etc., achieve simple and convenient use, short identification time, The effect of low production cost

Inactive Publication Date: 2011-06-22
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above two imported kits are more expensive, and because they contain 48 kinds of the same carbon sources, accounting for about 50% of the total carbon sources measured, and do not contain some common carbon sources, the results of the two kits are not comparable.

Method used

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  • Bacterium identification reagent kit as well as preparation method and uses thereof
  • Bacterium identification reagent kit as well as preparation method and uses thereof
  • Bacterium identification reagent kit as well as preparation method and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] (1) Activate the Enterobacter cloacae model strain Enterobacter cloacae ATCC 13047 with a medium plate for 1-2 times;

[0040](2), collect its fresh thallus, wash thalline 2 times with the centrifugation of 0.9% normal saline, wash with sterile water, dilute thalline, make OD 600 Bacterial suspension with a value of 1.5;

[0041] (3), take step 2 bacterial suspension 0.5ml, OD 600 =1.5, add 15ml of sterile semi-solid 0.2% agar-deionized water, shake well and mix completely;

[0042] (4) Use an eight-channel pipette to take 150 μL of the bacterial suspension in step 3 and add it to each well of the kit;

[0043] (5) Put the kit with the added bacterial suspension into a 37°C constant temperature bacterial incubator for cultivation, and determine the cultivation temperature according to the optimal growth temperature of the bacteria;

[0044] (6) After culturing for 20 hours, observe the kit, and use A1 as the control well to determine the utilization of bacterial carb...

Embodiment 2

[0047] Same as in Example 1, after activating the Rhizobium japonicum type strain NZP 2213 with LB plate, collect its fresh bacteria, wash with sterile water, and make bacterial suspension according to the above steps. Add 150 μL of bacterial suspension to each well of the kit with a pipette. Put the kit with the added bacterial suspension into a 37°C constant temperature bacterial incubator for cultivation, observe the kit after 4 days, and use A1 as the control well to determine the utilization of bacterial carbon sources. Observation results such as figure 2 shown. It can be seen that the rhizobia type strain NZP2213 can utilize dextrin, Tween 40, Tween 80, sucrose, D-glucose, D-mannose, D-galactose, L-fructose, D-fructose, D-xylose, xylose , D-ribose, D-trehalose, D-mannitol, L-arabinitol, D-sorbitol, hexyl alcohol, m-inositol and other 57 carbon sources, specifically attached figure 2 A2, A4, A5, A8, A9, A11, C1, C3, C4, C6, C7, C10, C11, B3, B5~7, B9, B11, B12, D1, ...

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Abstract

The invention discloses a bacteria identification kit, including indicator, culture medium and testing carbon sources. Thiazole blue is used as the indicator for indicating that whether bacteria can use the testing carbon sources. The invention is a kit which uses a 96-microtitor plate that is prearranged with the culture medium with 95 different carbon sources and is used for the rapid identification of carbon source utilization spectrum. One hole is not added with the carbon source for the comparison. The kit of the invention includes some carbon sources for detecting the specific physiological characteristics and common carbon sources. That using the thiazole blue as the indicator is convenient for naked eyes to observe that whether the bacteria grows and uses the carbon sources. And an eliasa can be used at the wavelength of 600nm to rapidly measure the growing condition of the bacteria. The kit of the invention has the advantages of low cost, short identification time, high sensitivity, convenient operation, accuracy, etc. the result can be made within 20-24 hours. And gram negative bacteria and positive bacteria can be identified.

Description

technical field [0001] The invention relates to the technical field of microbiological bacteria identification, in particular to a bacteria identification kit for identifying the utilization of the only carbon source of bacteria and a preparation method thereof. Background technique [0002] With the rise and development of the biological industry, rapid and accurate identification of microorganisms has become an urgent need in many fields such as medical clinics, environment, industry and agriculture. Conventional microbial identification techniques include observation of morphological structure and culture characteristics, physiological and biochemical tests, serum blood tests, rapid identification combinations, molecular biology methods, etc. These methods are either time-consuming, expensive, or difficult to master, and cannot meet the needs of a wider range of users. [0003] In the past 20 years, with the infiltration of advanced technologies such as microelectronics,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/04
Inventor 谭志远张武彭桂香蔡锦标
Owner SOUTH CHINA AGRI UNIV
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