Unlock instant, AI-driven research and patent intelligence for your innovation.

Probe, primer group and kit for detecting helicobacter pylori

A Helicobacter pylori and kit technology, applied in the biological field, can solve problems such as increasing medical burden, achieve low requirements, optimize reaction system and conditions, and save time

Pending Publication Date: 2021-11-16
山西康健恩生物科技有限公司
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Hp infection causes great suffering to patients and greatly increases the medical burden

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Probe, primer group and kit for detecting helicobacter pylori
  • Probe, primer group and kit for detecting helicobacter pylori
  • Probe, primer group and kit for detecting helicobacter pylori

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] The purpose of this example is to provide test materials.

[0039] 1.1 Standard strains and clinical samples

[0040] The ATCC Hp26695 standard strain was preserved by the department, and the gastric mucosa samples were from a hospital in Linfen.

[0041] 1.2 Primers and probes

[0042] Through the comparison and analysis of the known Helicobacter pylori nucleotide sequences in GenBank, a conserved region on ureI of Helicobacter pylori was found as the target gene. The screening of this gene was derived from the sequencing analysis of 80 strains from national sources by Linfen People's Hospital, and these strains represent the epidemic strains in China. Meanwhile, in this embodiment, human β-actin (β-actin) is selected as an internal reference gene. β-actin gene is often used as an internal reference gene for detecting pathogenic microorganisms from human specimens. The specific primer probe sequences used in this example are shown in Table 1, wherein the 5-end of u...

Embodiment 2

[0064] The purpose of this example is to optimize the multiplex real-time fluorescent quantitative PCR reaction system and reaction conditions of ureI and β-actin.

[0065] The Taq enzyme (5U / μl) of QIAGEN was selected as the reaction enzyme, and the optimal reaction system (25 μl) of the enzyme is shown in Table 2:

[0066] Table 2

[0067]

[0068]

[0069] Reaction conditions: 95°C for 5 minutes, 95°C for 15s, 60°C for 45s (collecting fluorescence), a total of 40 cycles.

[0070] Interpretation of results: samples with a Ct value less than or equal to 35 are positive results; samples with a Ct value greater than 35 are negative results; samples with a Ct value between 35 and 40 need to be repeated, and if the Ct value is still lower than 40 in repeated tests, it is judged as positive expansion. If it exceeds 40, it is judged as negative amplification.

Embodiment 3

[0072] The purpose of this example is to prepare a standard curve.

[0073] With the log value of the standard concentration template as the abscissa and the corresponding Ct value as the ordinate, the real-time fluorescence quantitative PCR standard curves of the Hp ureI and β-actin systems were drawn, respectively. The results showed that the amount of positive template was 1E 7 copies / μl~1×1E 3 In the range of copies / μl, the logarithmic value has a very good correlation with the Ct value (ureI R square is equal to 0.999, β-actin R square is equal to 0.998), and the amplification efficiency diagrams of ureI R and β-actin R are shown in figure 1 and 2 shown.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention provides a probe, a primer group and a kit for detecting helicobacter pylori, and relates to the technical field of biology. The probe for detecting the helicobacter pylori comprises a ureI probe, and the sequence of the ureI probe is shown as SEQ ID NO. 3. The primer group for detecting the helicobacter pylori comprises a ureI primer pair, the ureI primer pair comprises a ureIF with the sequence shown as SEQ ID NO.1 and a ureIR with the sequence shown as SEQ ID NO.2, and the primer group further comprises the probe. In addition, the invention further provides a kit containing the primer group, and the kit is used for detecting the helicobacter pylori in gastric mucosa tissues through multiple real-time fluorescent quantitative PCR. The primer group or the kit provided by the invention is high in detection sensitivity, good in specificity and low in requirements on detection tissues, the detection limit is 1E1copies / microliter, the detection accuracy reaches the quality level of the same product in the market, and the clinical test requirements can be met.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a probe, primer set and kit for detecting Helicobacter pylori. Background technique [0002] In 1983, Australian scholars Marshall and Warren first isolated Helicobacter pylroi (H. pylori or Hp) from gastric mucosa samples. Hp is a common Gram-negative microaerophilic bacillus that can colonize human gastric mucosa for a long time, and is closely related to many upper gastrointestinal diseases. More than half of the people in the world are infected with Hp, while the infection rate in our country is 55%. Hp infection has caused great suffering to patients and greatly increased the medical burden. WHO lists Hp as a biological carcinogen like hepatitis B and hepatitis C. Therefore, there is an urgent need for a method that can rapidly and specifically detect Hp infection. [0003] There are many methods for diagnosing Hp infection, including urease detection, bacterial culture, his...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/6851C12Q1/06C12N15/11C12R1/01
CPCC12Q1/689C12Q1/6851C12Q2600/16C12Q2600/166C12Q2531/113C12Q2537/143C12Q2561/101C12Q2545/101C12Q2545/113
Inventor 王青云吴玉庆赵琨
Owner 山西康健恩生物科技有限公司
Features
  • R&D
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com