Kit for detecting colorectal cancer indicator bacteria

A technology for colorectal cancer and indicator bacteria, which is applied in the field of kits for the detection of colorectal cancer indicator bacteria, can solve the problems of poor sensitivity, cumbersome pre-preparation, and difficulty in detecting colorectal cancer mutation signals, and achieve the effect of high sensitivity

Inactive Publication Date: 2020-01-03
SHANGHAI PASSION BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Not only that, traditional detection methods (such as fecal occult blood, digital examination, tumor markers, etc.) are not sensitive enough to detect mutation signals of early colorectal cancer, while colonoscopy detection requires high physical fitness of the tester, and the early The preparation is cumbersome and invasive, and the detection rate of colonoscopy in asymptomatic population is less than 1%

Method used

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  • Kit for detecting colorectal cancer indicator bacteria
  • Kit for detecting colorectal cancer indicator bacteria
  • Kit for detecting colorectal cancer indicator bacteria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0087] Example 1: SVM algorithm combined with indicator bacteria primer combination and fecal occult blood FIT index combination has higher detection accuracy than fecal occult blood FIT

[0088] 1 Detection of the abundance of 9 intestinal bacteria in feces and the results of fecal occult blood

[0089] 1.1 Sample source

[0090] A total of 1086 fecal samples from patients with colorectal cancer in the Anorectal Department of Liaoning Cancer Hospital and those who were healthy by colonoscopy were collected through cooperation.

[0091] 1.2 Specimen extraction

[0092] Use the feces collection box, tear off the adhesive, put it in a plastic bag; paste the box on the toilet, and excrete the feces into the feces collection box; take a sample (about 5 spoons of feces) into the feces storage tube with a sampling spoon, and then tighten the tube firmly Cover; put the feces preservation tube into the self-sealing tape, and store it at -80°C for later use.

[0093] The bacterial g...

Embodiment 2

[0150] Embodiment 2: Support vector machine method detection model parameter optimization

[0151] 1. Treat the feces

[0152] The specific experimental steps in this example are the same as those in Example 1. The feces of healthy people and patients with colorectal cancer were collected, the bacterial genome was extracted, PCR was performed with 16s rRNA primers, and the values ​​were recorded by the calculation method of the threshold line (Ct) of the amplification curve. Also perform fecal occult blood testing.

[0153] 2. Standardize the results of manure disposal

[0154] The specific result processing in this embodiment is the same as the steps in Embodiment 1. Standardized input reference set data file format, the first column is the sample number, the second to eighth columns are indicator bacteria 1, indicator bacteria 2, indicator bacteria 3, indicator bacteria 4, indicator bacteria 5, indicator bacteria 6, indicator bacteria 7 The ΔCt value, the ninth column is...

Embodiment 3

[0174] Embodiment 3: support vector machine method detects strain combination preferred 1

[0175] 1. Treat the feces

[0176] The specific experimental steps in this example are slightly different from those in Example 1. In this embodiment, 9 combinations of indicator bacteria were detected, and the performance of the 9 combinations of indicator bacteria and the 7 preferred indicator bacteria described in the present invention in identifying colorectal cancer were compared. The rest of the experimental steps are the same as in Example 1.

[0177] The feces of healthy people and colorectal cancer patients were collected in combination, the bacterial genome was extracted, PCR was performed with 16s rRNA primers, and the values ​​were recorded by the calculation method of the threshold line (Ct) of the amplification curve. Also perform fecal occult blood testing.

[0178] The primer sequences of 9 indicator bacteria are as follows:

[0179] Table 10

[0180]

[0181] ...

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Abstract

The invention discloses a kit for detecting colorectal cancer indicator bacteria. The kit is characterized by comprising the following components: a specific primer combination for identifying the indicator bacteria, wherein the primer combination is shown as SEQ ID NO. 1-14, and the indicator bacteria are the combination of fusobacterium nucleatum (F.nucleolus), anaerobic digestion streptococcus(P.anaerobicus), clostridium symbiosum (C.symbiolum), Porphyromonas saccharolytica (P.asaccharolytica), Prevotella intermedia (P.intermedia), Bacteroides fragilis (B.fragilis) and Streptococcus salivarius (S.salivarius). The kit for detecting colorectal cancer indicator bacteria disclosed by the invention has the beneficial effects that the colorectal cancer can be identified only by collecting astool sample. The principle is that the aim of identifying the colorectal cancer by detecting the colorectal cancer pathogenic bacteria in excrement, the sensitivity can reach 92.9%, and the specificity is 92.6%.

Description

technical field [0001] The invention belongs to the field of gene detection, in particular to a kit for detecting colorectal cancer indicator bacteria. Background technique [0002] Among the common types of human cancers, according to the statistics published in "Cancer incidence and mortality in China, 2014", the incidence rate of colorectal cancer is 17.2 / 105, ranking fifth in the incidence rate of tumors (the top four are lung cancer, breast cancer, and breast cancer respectively). cancer, stomach cancer, liver cancer). At the same time, colorectal cancer is also one of the most common causes of death. According to statistics, more than 600,000 people die from colorectal cancer every year. [0003] The occurrence of colorectal cancer involves progressive changes in intestinal flora structure, molecular genetics, and related histomorphological changes. It is a multifactorial process that interacts with internal factors and external factors such as the environment. It is...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/6886C12Q1/06C12Q1/14C12N15/11C12R1/01C12R1/145C12R1/46
CPCC12Q1/6886C12Q1/689
Inventor 孙子奎宣涛蔡庆乐
Owner SHANGHAI PASSION BIOTECHNOLOGY CO LTD
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