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Composition, kit and method for detecting human intestinal bacteria

A technology for detecting reagents and intestinal bacteria, applied in the directions of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of lack of immunity, affecting the health of the body, imbalance of intestinal flora, etc., to achieve specificity high effect

Active Publication Date: 2020-01-17
北京圣维尔医学检验实验室有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Under normal circumstances, the intestinal microbes in the human body form a relatively balanced state. Once the balance is disrupted, such as taking antibiotics, radiotherapy, chemotherapy, emotional depression, physical weakness, lack of immunity, etc., the intestinal flora will Loss of balance, which further affects the health of the body

Method used

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  • Composition, kit and method for detecting human intestinal bacteria
  • Composition, kit and method for detecting human intestinal bacteria
  • Composition, kit and method for detecting human intestinal bacteria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Embodiment 1, the method for detecting human intestinal bacteria

[0058] The detection sample of the present invention is feces, using the nucleic acid extraction / purification reagent based on the magnetic bead method (article number S1006, Hunan Shengxiang Biotechnology Co., Ltd., Xiangchang Machinery Equipment No. 20150021) to extract DNA, and perform the following operations in the sample processing room:

[0059] 2.1 Take soybean-sized feces into a 1.5mL sterilized centrifuge tube, add 1mL of normal saline to shake and mix, then centrifuge at 12,000rpm for 2 minutes, take 200μL of the upper layer solution for extraction, and add 300μL of DNA extraction solution 1 to the centrifuge tube;

[0060] 2.2 Add 200 μL of the sample to be tested in each tube; cover the tube cap, shake and mix for 10 seconds, and centrifuge instantaneously;

[0061] 2.3 Add 100 μL of DNA Extraction Solution 2-mix to each tube (mix well and then draw), shake and mix for 10 seconds, then let s...

Embodiment 2

[0075] Embodiment 2, the screening of primers and probes used in the present invention

[0076] Multiple sets of detection systems were designed for each type of bacteria, and the optimal system was screened for combined testing of joint detection. In the screening process, considering the economy, all probes used the FRET system of FAM-BHQ-1, and the primers and probes used were As shown in Table 1 below, the screening results of Bifidobacterium and Streptococcus were taken as an example to present the results.

[0077] Primers and probes designed in Table 1

[0078]

[0079]

[0080]

[0081] Wherein, S, R and Y are all degenerate bases, S=C / G, R=A / G, Y=C / T.

[0082] Wherein, the amplification system for detecting Bifidobacterium BIF is screened according to the method described in Example 1: as figure 1 As shown, the final selection curve is straight, and BIF-F, BIF-R and BIR-P without non-specific amplification are used as the final system for the detection of b...

Embodiment 3

[0085] Example 3, Amplification System Efficiency Detection of Each Reagent Combination

[0086] BIF is combined with BAC and STR to form a reagent combination, and the detection is performed according to the method described in Example 1. Compared with the BIF single strain amplification system, the efficiency of the amplification detection system is compared; LAC, BAC and BIF are combined to form a reagent combination , and detect according to the method as described in Example 1 and compare the efficiency of the amplification detection system with the LAC single bacterial species amplification system; Perform detection and compare the performance of the amplification detection system with the FAE, CLEP, and ER single-species amplification systems, and the results are as follows Figure 3-5 shown. It can be seen from the figure that Bifidobacterium BIF can be combined with Bacteroides BAC and Streptococcus STR in any combination. Any combination of Streptococcus BIF, combi...

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Abstract

The invention relates to the field of molecular biological detection, in particular to the field of human intestinal bacteria detection. The invention provides a composition for detecting seven intestinal bacteria, including Bifidobacterium (BIF), Streptococcus (STR), Bacteroides (BAC), Lactobacillus (LAC), Clostridium leptum (CLEP), Faecalibacterium (FAE) and Eubacterium rectale (ER), and also provides application of the composition in detecting human intestinal bacteria, a kit comprising the composition and a method for detecting human intestinal bacteria.

Description

technical field [0001] The present invention belongs to the field of molecular biology detection, specifically, belongs to the detection field of human intestinal bacteria; more specifically, the intestinal bacteria include Bifidobacterium (BIF), Streptococcus (STR), Bacteroides ( BAC), Lactobacillus (LAC), Clostridium tenebilis (CLEP), Fustridium prausaulius (FAE) and Eubacterium rectalum (ER) were seven kinds of intestinal bacteria. Background technique [0002] Intestinal microorganisms refer to the large number of microorganisms in the intestines of animals. This group of microorganisms lives in the intestines of animals and helps the host to complete a variety of physiological and biochemical functions. Under normal circumstances, the intestinal microbes in the human body form a relatively balanced state. Once the balance is disrupted, such as taking antibiotics, radiotherapy, chemotherapy, emotional depression, physical weakness, lack of immunity, etc., the intestinal ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/6851C12Q1/14C12Q1/06C12R1/46C12R1/145C12R1/01
CPCC12Q1/689C12Q1/6851C12Q2531/113C12Q2563/107Y02A50/30
Inventor 任小梅刘佳邓中平戴立忠
Owner 北京圣维尔医学检验实验室有限公司
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