Noninvasive helicobacter pylori gene detection kit and preparation and detection method thereof

A technology for detection of Helicobacter pylori and genes, applied in biochemical equipment and methods, measurement/inspection of microorganisms, DNA/RNA fragments, etc., can solve the problems of manpower and material resources and detection time without cost saving, and achieve the preparation method And the effect of simple operation steps, high sensitivity, improved sensitivity and specificity

Active Publication Date: 2014-06-04
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 o

Method used

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  • Noninvasive helicobacter pylori gene detection kit and preparation and detection method thereof
  • Noninvasive helicobacter pylori gene detection kit and preparation and detection method thereof
  • Noninvasive helicobacter pylori gene detection kit and preparation and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1 The design and specific test of the primer

[0052] The 16S RNA of the Helicobacter pylori is the target sequence. The design primer is as follows:

[0053] The primer is 5' -ttgggataGCCATTGGAACG -3 '

[0054] 3' -gttccgataCTGCCCATAGC -5 ';

[0055] Breitry ---- Helicobacter pylori Sydney Helicobacter Pylori SS1 (referred to as HP) (purchased from Hebei Medical University, from the Institute of Popular Diseases of the China Disease Prevention and Control Center) as a target sequence.The product is between 100-200bp, and the alkali base of the primer is randomly distributed and uniform. The primer itself cannot form a di cluster. The length of the primer is between 17-25 bases.The content is 45-55%, the primers are not rich in AT and GC, there is no T / C, A / G continuous structure (2-3), and the primer 3 ′ is not on the third place of the password, and the 16S RNA is 16S RNAFor the conservative sequence of Helicobacter pylori, all the Helicobacter pylori can be detec...

Embodiment 2

[0058] Example of Example 2 Primers of Primers

[0059] The above primers 5' -ttgggataGCCATTGGAACG -3 '

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

[0061] The primer melted chain 8-10s at 90-98 ° C; then cooled to 56-66 ℃ annealing and extending 25-30s;

[0062] Two steps cycle 30Cycle.The existing process is to reduce the temperature to about 55 ° C for a period of time, and then increase the temperature to about 72 ° C, and the annealing extension of the present invention is a step, because the annealing extension adopted by the present invention is performed under the same temperature conditions under the same temperature conditions.Therefore, there is no need to cool the smelting product after two cooling, but directly cool down the product required when the product is extended to obtain the primer amplification fragment. Compared with the PCR amplification response of the existing primer, it eliminates a cooling step to cool down the cooling ...

Embodiment 3

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

[0066] A kit of a non -invasive nucleic acid detection of Helicobacter pylori includes R1 reagent: primer 5' -ttgggatagcccattgaaacg -3 '

[0067] 3' -gttccgataCTGCCCATAGC -5 ',

[0068] R2 reagent: SYBR Green TAQ MIX (CW0078, CWBIO), other PCR TAQ enzyme pre -mixing liquids are also available; R3 reagent: sterile distilled water (201305, homemade).

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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 present invention involves a primer, kit and its preparation and detection methods of non -invasive pylori gene detection. It is designed to design a primer to configure a kit, which provides a non -invasive, simple, fast, and sensitive Helicobacter pylori geneThe detection kit and detection method provide methods and reagents for detecting people and animal Helicobacter pylori infections. Background technique [0002] Pepsitobaus ( Helicobacter Pylori Referred to as HP) was discovered by Warren and Warshail in Australia and one of the pathogens that have been recognized as the most common chronic infectious diseases.At this stage, clinical testing HP is mainly divided into two categories: invasiveness and non -invasiveness. Invasive testing is generally gastroscopy and serum antibody detection; non -invasive testing is a theifx of the isotope C marking the respiratory experiment and the hemapacorcoticobacter pylori antigen detection.Although these dete...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04C12N15/11
CPCC12Q1/686C12Q2531/113
Inventor 祝洁张勇
Owner SICHUAN VACCINE TECH
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