A probe for absolute quantification of aspergillus tubingensis and its application

A probe and signal probe technology, applied in the biological field, can solve the problems of low species resolution, insufficient detection accuracy, cumbersome operation of quantitative methods, etc., and achieve the effect of high accuracy and improved detection limit

Active Publication Date: 2022-04-29
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] A kind of probe, test kit and application for the absolute quantification of Aspergillus tubingensis of the present invention solve at least one technical problem as follows: (1) the existing method cannot realize the total amount detection of all Aspergillus tubingensis; (2) now Some 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 for absolute quantification of aspergillus tubingensis and its application
  • A probe for absolute quantification of aspergillus tubingensis and its application
  • A probe for absolute quantification of aspergillus tubingensis and its application

Examples

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Effect test

Embodiment 1

[0059] Embodiment 1: Aspergillus tubingensis quantitative probe combination reagent

[0060] 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 .

[0061] 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

[0062] Embodiment 2: Aspergillus tubingensis quantitative kit and its use

[0063] The Aspergillus tubingensis quantitative 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.

[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 method of use is:

[0066](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.

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

Embodiment 3

[0073] Embodiment 3: Aspergillus tubingensis quantitative kit

[0074] Aspergillus tubingensis quantitative kit, containing 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 .

[0075] 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 probe for absolute quantification of Aspergillus tubingensis and an application thereof, belonging to the fields of biology, fermentation and detection. The Aspergillus tubingensis quantitative probe and kit of the present invention can realize the detection of the total amount of Aspergillus tubingensis, do not need to use expensive instruments for the detection and quantification of Aspergillus tubingensis, 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 Aspergillus tubingensis quantification, and have the characteristics of fast, convenient, cheap and accurate.

Description

technical field [0001] The invention relates to a probe for absolute quantification of Aspergillus tubingensis and an application thereof, belonging to the fields of biology, fermentation and detection. Background technique [0002] Aspergillus tubingensis widely exists in traditional fermented foods, and participates in the biological reaction process in the fermentation process, including liquor, Puer tea, etc. For example, during the fermentation process of liquor, Aspergillus tubingensis can metabolize and produce acid compounds to inhibit the growth of bacteria, and at the same time secrete a variety of enzymes and have strong stress resistance. Therefore, it is necessary to track the biomass of Aspergillus tubingensis in real time, which has important guiding significance for judging the stability of fermentation batches and regulating fermentation parameters. However, at present, most traditional fermented food systems are multi-strain co-fermentation systems, and th...

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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