Helicobacter pylori specific molecular marker and detection kit

A technology of Helicobacter pylori and detection kit, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, microorganisms, etc. It can solve the problems of insufficient sensitivity and achieve the effect of improving sensitivity, high sensitivity and strong specificity

Active Publication Date: 2019-03-29
PEKING UNIV THIRD HOSPITAL
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a specific molecular marker and detection kit for gastric Helicobacter pylori, and realize the specific detection of Helicobacter pylori according to the principle of PCR-reverse dot hybridization method, so as to solve the problem of detection of Helicobacter pylori in the prior art. The invasive detection of Helicobacter is not suitable for children and the elderly, and the sensitivity is not enough. The molecular marker and kit of the present invention can also be used as a positive reference in the detection of other Helicobacter pylori genes such as cag, vec and other drug-resistant genes. In order to monitor all the processes of other genes in genome extraction, PCR and hybridization, and promote further in-depth research on Helicobacter pylori

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  • Helicobacter pylori specific molecular marker and detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Embodiment 1: Gastric Helicobacter pylori detection kit composition

[0047] The kit of the present invention consists of the following:

[0048]

[0049] Table 1 The quantity and main components of each component in the Helicobacter pylori detection kit

[0050]

[0051]

[0052] PCR amplification primer sequence in table 2 Helicobacter pylori detection kit

[0053]

[0054]

[0055] Detection probe sequence in table 3 Helicobacter pylori detection kit

Embodiment 2

[0056] Embodiment 2: detection method of gastric Helicobacter pylori

[0057] The kit of the present invention can use gastric mucosal tissue and feces as samples. Correspondingly, the following requirements should be paid attention to when sampling. When gastric mucosal tissue is used as a sample: during gastroscopy, the gastric mucosa of the patient's lesion is taken, sealed and sent for examination. Due to the differences in the colonization sites of Helicobacter pylori, in order to increase the detection rate, it is recommended to sample multiple parts of the gastric mucosa of each patient, among which gastric antrum mucosa is the most common. When used as a sample: adjust the patient's diet and reduce the intake of plant foods to reduce the polysaccharides in the stool sample, thereby avoiding PCR inhibition.

[0058] 【Testing method】

[0059] 1. Extraction of DNA

[0060] Genomic DNA extraction can be performed according to the instructions of commercial DNA extractio...

Embodiment 3

[0088] Example 3: Detection of gastric Helicobacter pylori infection in patients

[0089] To ensure the reliability of the test results, 10 gastric antrum mucosa samples from Helicobacter pylori positive patients and 10 gastric antrum mucosa samples from Helicobacter pylori negative patients were selected for follow-up detection.

[0090] Histological section staining is a good method for clinicians to conduct clinical diagnosis. The analysis of the staining results can clearly determine whether there is Helicobacter pylori infection in gastric tissue. The tissue with Helicobacter pylori infection is positive, and the tissue without infection is negative. . In this embodiment, the histological section staining method was selected as a control.

[0091] Adopt the detection method of embodiment 2 respectively to above-mentioned samples, use this specific gastric Helicobacter pylori detection kit to detect, all positive samples of the result are as follows: figure 2 As shown i...

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Abstract

The invention relates to a helicobacter pylori specific molecular marker and a detection kit thereof. The kit is used for performing specific detection on helicobacter pylori in samples of gastric mucosa, gastric juice, excrement and throat swab according to PCR-reverse dot blot. A target gene at which the helicobacter pylori specific molecular marker aims is a urease ureH gene. With the adoptionof the kit, sensitivity of clinical examination of helicobacter pylori is improved greatly, and furthermore, helicobacter pylori infected persons, especially old people and children can be subjected to noninvasive screening, so that early detection and early treatment can be realized; besides, for detection of other helicobacter pylori genes such as cag and vec and other drug resistance genes, thekit can be taken as positive reference and used for monitoring all processes such as genome extraction, PCR and hybridization of other genes.

Description

technical field [0001] The invention belongs to the field of biological detection, and in particular relates to a gastric Helicobacter pylori-specific molecular marker and a detection kit, in particular to a kit for detecting gastric Helicobacter pylori by PCR-reverse dot hybridization. Background technique [0002] my country is the hardest-hit area for Helicobacter pylori (Hp) infection, with an infection rate as high as 40%-60%. The common meal system and the eating habits and poor sanitation environment in a few underdeveloped areas make the infection rate of children reach 30%-40%. Most adult H. pylori infections are acquired during childhood, mostly in early childhood, and are generally difficult to clear spontaneously after infection, leading to lifelong infection. Therefore, the screening and treatment research of Hp is very important to reduce the infection rate of Hp and improve the cure rate of Hp. [0003] At present, there are mainly the following methods for H...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/6834C12Q1/04C12N15/11C12R1/01
CPCC12Q1/6834C12Q1/689C12Q2531/113C12Q2545/113
Inventor 宋志强周丽雅
Owner PEKING UNIV THIRD HOSPITAL
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