Fungus absolute quantification kit and application thereof

A detection kit and fungi technology, applied in the biological field, can solve the problems of inappropriate detection in time, complicated operation of quantitative methods, and inability to detect the total amount of fungal microorganisms.

Active Publication Date: 2021-03-09
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] A method, kit and application for the absolute quantification of fungal microorganisms 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 fungal microorganisms; (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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  • Fungus absolute quantification kit and application thereof
  • Fungus absolute quantification kit and application thereof
  • Fungus absolute quantification kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Embodiment 1: fungal microorganism quantitative probe combination reagent

[0061] Probe combination reagent; containing independently packaged signal probe reagent and quenching probe reagent; wherein, the signal probe sequence is as shown in SEQ ID NO.1, and the quenching probe sequence is as shown in SEQ ID NO.2 .

[0062] Signal probe reagents and quenching probe reagents are in dry powder or liquid form; in dry powder form, they can be diluted to an appropriate concentration before the experiment, for example, diluted with sterile water or buffer solution to a concentration of 20 μM; in liquid form , the concentration can be 20-200μM, and the reagent can be diluted before use, or used directly.

Embodiment 2

[0063] Embodiment 2: fungal microorganism quantification kit and its use

[0064] The fungal microorganism quantification kit contains independently packaged signal probe reagents and quenching probe reagents; wherein, the signal probe sequence is shown in SEQ ID NO.1, and the quenching probe sequence is SEQ ID NO.2.

[0065] 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.

[0066] The method of use is:

[0067] (1) Solution configuration. Configure 100nM heme solution (reagent 1); configure 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 2,2-azino-bis-( 3-Ethylbenzodihydrothiazoline-6-sulfonic acid) diammonium salt (ABTS) (reagent 3) and 7mM H 2 o 2 Solution (reagent 4); solvents are sterile water.

[0068] (2) The signal probe forms a double strand with the sample DNA. Add 4...

Embodiment 3

[0074] Embodiment 3: fungal microorganism quantification kit

[0075] The fungal microorganism quantification kit contains independently packaged signal probe reagents and quenching probe reagents; wherein, the signal probe sequence is shown in SEQ ID NO.1, and the quenching probe sequence is shown in SEQ ID NO.2 .

[0076] The kit also contains 100nM heme solution (reagent 1), Tris-HCL buffer, 7mM 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 fungus absolute quantification kit and application thereof, and belongs to the fields of biology, fermentation and detection. A fungal microorganism quantitative probe and the kit of the invention can realize detection of the total amount of all fungal microorganisms, do not need to use expensive instruments when used for detection and quantification of fungal microorganisms, and can quickly complete quantification work within 2.5 hours. Meanwhile, a sample used does not need to be subjected to nucleic acid extraction. The probe and detection kit can be used for quantification of fungal microorganisms, and have characteristics of quickness, convenience, cheapness and accuracy.

Description

technical field [0001] The invention relates to a kit for absolute quantification of fungi and an application thereof, belonging to the fields of biology, fermentation and detection. Background technique [0002] There are complex microbial flora involved in the brewing process of traditional fermented food, among which fungi are important functional microorganisms. These microorganisms are closely related to the quality of the final product. During the fermentation process, fungal microorganisms participate in a series of physiological and biochemical reactions related to substrate degradation and product synthesis, and convert available glycogen, protein and other macromolecules in raw materials into alcohols , aldehydes, acids, esters and other flavor components, endowing the product with unique flavor and sensory characteristics. Therefore, real-time tracking of the growth trend of fungal microorganisms during the fermentation process has important guiding significance ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6816C12Q1/06C12N15/11
CPCC12Q1/6895C12Q1/6816C12Q2521/337C12Q2525/301Y02A50/30
Inventor 吴群徐岩杜如冰
Owner JIANGNAN UNIV
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