A probe, method and application for absolute quantification of Zygomyces bayeri

A technology of Bayer Zygomycetes and probes, which is applied in biochemical equipment and methods, measuring devices, and microbial determination/inspection, etc. It can solve the problems of low species resolution, insufficient detection accuracy, and high-cost instruments and equipment of quantitative methods. and other problems, to achieve the effect of improved detection limit and high accuracy

Active Publication Date: 2022-04-15
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] A kind of probe, kit and application for the absolute quantification of Zygosaccharomyces bailii of the present invention solve at least one of the following technical problems: (1) the existing method cannot realize the total amount detection of all Zygosaccharomyces bailii; (2) the existing Quantitative methods have low species resolution and / or insufficient detection accuracy; (3) Existing quantitative methods require high-cost instruments and / or strict operating environments, and are not suitable for timely detection after production sampling; (4) Existing quantitative methods are cumbersome to operate, etc.

Method used

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  • A probe, method and application for absolute quantification of Zygomyces bayeri
  • A probe, method and application for absolute quantification of Zygomyces bayeri
  • A probe, method and application for absolute quantification of Zygomyces bayeri

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Example 1: Zygosaccharomyces bailii quantitative probe combination reagent

[0060] Probe combination reagent; contains independently packaged signal probe reagent and quenching probe reagent; wherein, the signal probe sequence is shown in SEQ ID NO.1, and the quenching probe sequence is shown in SEQ ID NO.2 .

[0061] Signal probe reagents and quench probe reagents, in dry powder or liquid form; in dry powder form, they can be diluted to an appropriate concentration before the experiment, for example, use sterile water or buffer to dilute to a concentration of 20 μM; in liquid form , the concentration can be 20-200μM, the reagent can be diluted before use, or used directly.

Embodiment 2

[0062] Example 2: Zygosaccharomyces bailii quantitative kit and its use

[0063] The Zygosaccharomyces bailii quantitative kit contains independently packaged signal probe reagents and quench probe reagents; wherein, the signal probe sequence is shown in SEQ ID NO.1, and the quench probe sequence is shown in SEQ ID NO.2.

[0064] When the kit is used, it can be mixed with heme, buffer, 2,2-azino-bis-(3-ethylbenzodihydrothiazoline-6-sulfonic acid) diammonium salt (ABTS), H 2 O 2 With the use of.

[0065] The usage method is:

[0066](1) Solution configuration. Prepare 100nM heme solution (reagent 1); prepare Tris-HCL with a final concentration of 50mM, KCl with a final concentration of 50mM, and a final pH of 7.9 (reagent 2); 7mM of 2,2-azino-bis-( 3-ethylbenzodihydrothiazoline-6-sulfonic acid) diammonium salt (ABTS) (reagent 3) and 7 mM H 2 O 2 Solution (reagent 4); all solvents are sterile water.

[0067] (2) The signal probe forms a double strand with the sample DNA. ...

Embodiment 3

[0073] Example 3: Zygosaccharomyces bailii quantification kit

[0074] Zygosaccharomyces bailii quantitative kit, containing independently packaged signal probe reagents and quench probe reagents; the signal probe sequence is shown in SEQ ID NO.1, and the quench probe sequence is shown in SEQ ID NO.2 .

[0075] The kit also contains 100 nM heme solution (reagent 1), Tris-HCL buffer, 7 mM 2,2-azino-bis-(3-ethylbenzodihydrothiazoline-6-sulfonic acid) ) diammonium salt (ABTS), 7mM H 2 O 2 solution.

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Abstract

The invention discloses a probe, method and application for absolute quantification of Zygomyces zygosacrum Bayer, and belongs to the fields of biology, fermentation and detection. The Zygomycetes quantitative probe and kit of the present invention can realize the detection of the total amount of Zygomycetes Bayer, do not need to use expensive instruments when used for detection and quantification of Zygomycete Bayer, and can quickly complete the quantitative work within 2.5 hours. At the same time, the samples used in the present invention do not necessarily have to undergo nucleic acid extraction. The probe and detection kit based on the present invention are used for the quantification of Zygomycetes Bayer, and have the characteristics of rapidity, convenience, cheapness and accuracy.

Description

technical field [0001] The invention relates to a probe, method and application for absolute quantification of Zygosaccharomyces bayer, belonging to the fields of biology, fermentation and detection. Background technique [0002] Zygosaccharomyces bailii is a common yeast in a variety of fermented food systems, such as red wine, tea, white wine and vinegar, and plays an important role in the fermentation process. For example, in wine research, Zygosaccharomyces bailii and The co-cultivation of Saccharomyces cerevisiae can increase the overall ethyl acetate content, and it has a strong flavor metabolism. Studies have shown that Zygosaccharomyces bailii can metabolize to produce 33 important volatile compounds, so the biomass of Zygosaccharomyces bailii can be tracked in real time. It has important guiding significance for judging the stability of fermentation batches and regulating fermentation parameters. However, most of the traditional fermented food systems are multi-str...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/30
CPCG01N21/6428C12Q1/30G01N2021/6432
Inventor 吴群徐岩杜如冰
Owner JIANGNAN UNIV
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