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Preparation method of reagent for quantitatively determining Helicobacter pylori antigen in excrement

A technology for Helicobacter pylori and anti-Helicobacter pylori is applied in the field of medical testing, which can solve the problems of application limitation, cannot truly reflect the Helicobacter pylori in the stomach, etc., and achieves the effects of no potential safety hazard, not easy to interfere, and high detection efficiency.

Inactive Publication Date: 2019-02-15
SHENZHEN HONGMED INFAGEN CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The existing methods for serum antibody detection include enzyme-linked immunoassay, western blot, colloidal gold method, etc., but since the human serum still contains the corresponding antibody after the eradication of Helicobacter pylori, the presence of Helicobacter pylori infection can be determined by serum antibody Specificity problem, cannot truly reflect whether there is still Helicobacter pylori in the stomach
13C / 14C breath test, as a method for detecting Helicobacter pylori infection, is widely used clinically, but there are radioactive problems and special equipment is required, so the application in families and social health centers is greatly limited

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  • Preparation method of reagent for quantitatively determining Helicobacter pylori antigen in excrement
  • Preparation method of reagent for quantitatively determining Helicobacter pylori antigen in excrement
  • Preparation method of reagent for quantitatively determining Helicobacter pylori antigen in excrement

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Embodiment 1

[0035] This implementation provides a method for preparing a reagent for the quantitative determination of Helicobacter pylori antigen in feces, the reagent includes R1 reagent and R2 reagent, wherein, in terms of mass percentage, the R1 reagent contains 0.1-10% electrolyte, 0.1-10% stabilizer, 0.1-10% surfactant, 0.1-5% preservative, and the balance is 1-100mmol / L MES buffer.

[0036] 1. Preparation of R2 reagent

[0037] The R2 reagent includes latex particles coated with anti-Helicobacter pylori antigen (urease) polyclonal antibody, electrolyte, stabilizer, surfactant, preservative and buffer, and the R2 reagent is prepared by the following process:

[0038] S1. Select any protein subunit of urease as the Helicobacter pylori antigen. Urease is composed of two protein subunits, namely UreA and UreB. Cloning the UreA or UreB gene into the pGEX-4T-1 plasmid, The cloning sites are BamHI and EcoRI in the pGEX-4T-1 plasmid (such as figure 1shown), the pGEX-4T-1 plasmid contains...

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Abstract

The invention discloses a preparation method of a reagent for quantitatively determining a Helicobacter pylori antigen in excrement. The method comprises following steps of preparing an R2 reagent: S1, extracting Helicobacter pylori antigen (urease) protein, cloning a urease gene into a pGEX-4T-1 plasmid, fermenting and expressing by escherichia coli, and purifying by affinity chromatography to obtain a recombinant Helicobacter pylori antigen (urease); S2, preparing a polyclonal antibody of an anti-Helicobacter pylori antigen (urease); S3, preparing latex particles coated with the polyclonal antibody of the anti-Helicobacter pylori antigen (urease); and S4, dispersing in buffer solution to obtain the R2 reagent. The method is a latex enhanced immunoturbidimetry method, the prepared reagentis solution capable of directly detecting Helicobacter pylori through excrement, detection can be completed by only a excrement sample, and therefore sampling is convenient. The human body is free ofpotential safety hazards, detection efficiency is high, stability is good, specificity is high, and probability of interference is low.

Description

technical field [0001] The invention belongs to the technical field of medical detection methods, and relates to a method for rapid quantitative determination of Helicobacter pylori antigen (urease) in feces, in particular to a reagent for rapid quantitative determination of Helicobacter pylori antigen in feces by latex immune turbidimetry method of preparation. Background technique [0002] A large number of studies have shown that Helicobacter pylori (Hp) infection is an important cause of chronic gastritis and peptic ulcer, and is closely related to gastric cancer and gastric mucosa-associated lymphoid tissue lymphoma. At present, there are many methods to evaluate Hp infection clinically, which are mainly divided into two categories: invasive and non-invasive detection methods. [0003] Among them, the invasive detection method is to use endoscope to obtain gastric biopsy tissue for detection, generally including urease test method, pathological detection method and bac...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/531
CPCG01N33/56911G01N33/531
Inventor 侯志波张伟温建超翁琪墁
Owner SHENZHEN HONGMED INFAGEN CO LTD
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