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Primers, probes and kit for helicobacter pylori infection and typing detection

A Helicobacter pylori, primer probe technology, applied in the field of molecular biology, can solve the problems of non-invasive, difficult Helicobacter pylori detection reagents, detection sensitivity, and low specificity, and achieve strong pertinence, good promotion and application prospects, and reduce patient pain effect

Pending Publication Date: 2022-02-08
XIAMEN SCIENDOX BIOLOGICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] In order to solve the problem that the existing Helicobacter pylori infection and typing detection reagents are difficult to achieve complete non-invasiveness, and the detection sensitivity and specificity are not high, the present invention provides a primer set, which includes:

Method used

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  • Primers, probes and kit for helicobacter pylori infection and typing detection
  • Primers, probes and kit for helicobacter pylori infection and typing detection
  • Primers, probes and kit for helicobacter pylori infection and typing detection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Embodiment 1, the design and synthesis of primers and probes of the present invention

[0047] Specific primers and probes were designed using Primer Premier 3.0 and BeaconDesigner software for the specific sites of the genes UreA and CagA in the human genome and the internal reference gene ACTB (see the sequence of the human genome published in the NCBI database). Primers and probes were synthesized by Sangon Bioengineering (Shanghai) Co., Ltd.

[0048] The present invention simultaneously designs and verifies multiple pairs of primers and probe combination sequences for the UreA and CagA target gene fragment nucleic acid sequences related to Helicobacter pylori infection and typing. After repeated optimization experiments (see Experimental Example 1), the following are optimized (Table 1) A set of detection gene UreA, CagA fragment primer probe sequence is the optimal sequence.

[0049] Table 1 Detection of gene-specific region detection specific primer probe sequenc...

Embodiment 2

[0052] Embodiment 2, the reagent composition of Helicobacter pylori infection and typing detection of the present invention

[0053] Including UreA forward primer (primer 3), UreA reverse primer (primer 4), UreA detection probe (probe 2); CagA forward primer (primer 5), CagA reverse primer (primer 6), CagA detection probe Needle (probe 3); internal reference gene ACTB forward primer (primer 1), ACTB reverse primer (primer 2), ACTB detection probe (probe 1), PCR buffer, dNTPs, MgCl 2 .

[0054] The dosage ratio is shown in Table 2:

[0055] Table 2 PCR reagent composition

[0056]

Embodiment 3

[0057] Embodiment 3, the kit of Helicobacter pylori infection and typing detection of the present invention

[0058] The dosage ratio is shown in Table 3.

[0059] Table 3 Composition of the kit

[0060]

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Abstract

The invention provides high-specificity primers and probes aiming at target genes UreA and CagA. The sequences of the primer and probe group are as shown in SEQ ID NO.1-6. A kit for detecting helicobacter pylori infection and typing on the basis of real-time fluorescent quantitative PCR is prepared on the basis of the primer probe group, the helicobacter pylori can be conveniently and quickly detected and typed on the molecular level, the detection efficiency is high, the pertinence is strong, and the result is accurate and reliable; DNA extracted from excrement and saliva can be directly adopted for detection, high accuracy can be guaranteed while complete noninvasive is achieved, pain of a patient is relieved, the acceptability of the patient is improved, and the method has very good application and popularization prospects.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to a primer, a probe and a reagent kit for the infection and typing detection of Helicobacter pylori. Background technique [0002] Helicobacter pylori (Hp) is a spiral-shaped, slightly anaerobic, Gram-negative bacillus, which has a strong survival ability and can survive in the strong acidic environment in the stomach. Epidemiological studies have shown that the infection rate of Helicobacter pylori (Hp) is very high, and about 50% of the people in the world are infected. The incidence rate is related to factors such as socioeconomic level, population density, public health conditions, and water supply. Studies have shown that Hp infection mostly occurs in childhood, and the genotypes of strains infected by children are not changed by environmental migration, and are often consistent with the Hp genotypes infected by parents. In addition, Hp infection is also the most...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/689C12Q1/6851C12Q1/04
CPCC12Q1/689C12Q1/6851C12Q2600/166C12Q2531/113C12Q2563/107C12Q2545/113C12Q2545/101C12Q2561/113
Inventor 杨璐平颜宏利于修安宋立华蔡海芳
Owner XIAMEN SCIENDOX BIOLOGICAL TECH CO LTD