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A probe and kit for absolute quantification of lactobacillus jinshani

A technology for detection kits and probes, which is applied in the direction of measuring devices, microbial determination/inspection, instruments, etc., can solve the problems of quantitative methods such as high-cost instruments and equipment, cumbersome quantitative methods, and insufficient detection accuracy

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, kit and application for the absolute quantification of Lactobacillus jinshani of the present invention solve at least one technical problem as follows: (1) the existing method cannot realize the total amount detection of all Lactobacillus jinshani; (2) the present invention 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 and kit for absolute quantification of lactobacillus jinshani
  • A probe and kit for absolute quantification of lactobacillus jinshani
  • A probe and kit for absolute quantification of lactobacillus jinshani

Examples

Experimental program
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Effect test

Embodiment 1

[0060] Embodiment 1: Lactobacillus jinshani 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: Lactobacillus jinshani quantitative kit and its use

[0064] The Lactobacillus jinshani 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.

[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 H2 o 2 Solution (reagent 4); solvents are sterile water.

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

Embodiment 3

[0074] Embodiment 3: Lactobacillus jinshani quantitative kit

[0075] Lactobacillus jinshani 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 .

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

Description

technical field [0001] The invention relates to a probe and kit for absolute quantification of Lactobacillus jinshani, belonging to the fields of biology, fermentation and detection. Background technique [0002] Lactobacillus jinshani is widely distributed in the traditional fermented food brewing system. For example, in the liquor brewing system, the abundance of Lactobacillus jinshani increased sharply after 5 days of fermentation, and became the dominant microorganism in the middle and late stages of fermentation. Through correlation analysis, it was found that this bacterium and other microorganisms showed Significant correlation, which plays an important role in the fermentation process; it is also reported to be an important functional microorganism in the vinegar fermentation system. The biomass and succession of the bacteria in the food fermentation process are important microorganisms that characterize the normal fermentation of food. Therefore, tracking the bioma...

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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