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A kind of non-invasive helicobacter pylori gene detection kit and its preparation and detection method

A technology for Helicobacter pylori and gene detection, which is applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve the problems of human and material resources and detection time without cost saving, and achieve economical operation. The process and operation time, preparation method and operation steps are simple, and the specificity is good.

Active Publication Date: 2016-05-18
SICHUAN VACCINE TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the existing Hp detection methods are usually invasive. For example, the examination of tissue section staining is to take gastric mucosal specimen slices or smear staining microscope to examine Hp. Another example is the immunohistochemical staining of tissue sections. Staining is used to detect Hp, and generally it cannot be used as a routine diagnostic method. The media for these tests need to go through a complicated collection and production process. Material resources and inspection time are not cost-effective

Method used

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  • A kind of non-invasive helicobacter pylori gene detection kit and its preparation and detection method
  • A kind of non-invasive helicobacter pylori gene detection kit and its preparation and detection method
  • A kind of non-invasive helicobacter pylori gene detection kit and its preparation and detection method

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Effect test

Embodiment 1

[0051] Design and specificity test of embodiment 1 primer

[0052] Using the 16sRNA of Helicobacter pylori as the target sequence, the primers were designed as follows:

[0053] Primer is 5'-TTGGGATAGCCATTGGAAACG-3'

[0054] 3'-GTTCCGATACTGCCCATAGGC-5';

[0055] Using the 16sRNA of the strain---Helicobacter pylori Sydney strain Helicobacter pyloriSS1 (referred to as Hp) (purchased from Hebei Medical University, from the Institute of Epidemiology, Chinese Center for Disease Control and Prevention) as the target sequence, the primers obtained were located in the coding region 5 In the 300-400bp region at the 'end, the amplification product is between 100-200bp, the bases of the primers are randomly distributed and uniform, the primers themselves cannot form dimers, and the length of the primers is between 17-25 bases. The difference cannot be 3 bases, the G+C content is 45-55%, the primer is not in the area rich in AT and GC, there is no T / C, A / G continuous structure (2-3), an...

Embodiment 2

[0058] The amplification reaction of embodiment 2 primers

[0059] The above primer 5'-TTGGGATAGCCATTGGAAACG-3'

[0060] The PCR amplification reaction of 3'-GTTCCGATACTGCCCATAGGC-5' is:

[0061] Melt the primer at 90-98°C for 8-10s; then cool down to 56-66°C for annealing and extension for 25-30s;

[0062] Two-step cycle 30cycle. The existing process is to lower the temperature to about 55°C for annealing treatment for a period of time, and then raise the temperature to about 72°C for extension, but the annealing extension of the present invention is a step, because the annealing extension adopted in the present invention is carried out under the same temperature condition Therefore, it is not necessary to cool the smelted product twice, but directly cool the product to the temperature required for extension to obtain the primer amplified fragment. Compared with the PCR amplification reaction of the existing primer, one cooling step is omitted , which saves the operation p...

Embodiment 3

[0065] Example 3 A kind of non-invasive Helicobacter pylori gene detection kit

[0066] A kit for non-invasive nucleic acid detection of Helicobacter pylori including R1 reagent: primer 5'-TTGGGATAGCCATTGGAAACG-3'

[0067] 3'-GTTCCGATACTGCCCATAGGC-5',

[0068] R2 reagent: SYBRGreenTaqMIX (CW0078, CWBio), other PCRTaq enzyme premixes are also available; R3 reagent: sterile distilled water (201305, self-made).

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Abstract

The invention discloses a noninvasive helicobacter pylori gene detection primer and kit and a preparation and detection method thereof. The kit comprises a specific primer, a PCR (Polymerase Chain Reaction) amplification method with simple steps is adopted, the sample is subjected to noninvasive collection and manufacture, and pain of patients in the detection process is lightened. The defects in the prior art are overcome; the invention provides a simple, rapid and sensitive noninvasive helicobacter pylori gene detection kit and a preparation and detection method thereof. The method is simple in steps, the kit is a kit for sensitively detecting the helicobacter pylori, invasive sampling by using gastroscope and the like is avoided, the pain of the patients during detection can be reduced, and the time and cost of manpower and material resources are greatly reduced.

Description

technical field [0001] The invention relates to a primer, a kit for non-invasive Helicobacter pylori gene detection and a preparation and detection method thereof. A primer is specifically designed and configured into a kit to provide a non-invasive, simple, fast and sensitive Helicobacter pylori gene The detection kit and detection method provide methods and reagents for detecting Helicobacter pylori infection in humans and animals. Background technique [0002] Helicobacter pylori (Hp for short) was discovered by Warren and Warshail in Australia in 1983, and has been recognized as one of the most common pathogens of chronic infectious diseases. At present, clinical detection of Hp is mainly divided into two categories: invasive and non-invasive. Invasive detection is generally gastroscopy detection and serum antibody detection; non-invasive detection is isotope C-labeled breath test and fecal Helicobacter pylori antigen detection. Although these detection methods are rout...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12Q1/04C12N15/11
CPCC12Q1/686C12Q2531/113
Inventor 祝洁张勇
Owner SICHUAN VACCINE TECH
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