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Tjn molecular kit for diagnosing virulent strains of helicobacter pylori

A Helicobacter pylori and kit technology, applied in the field of molecular diagnostic kits used to detect virulent strains of Helicobacter pylori, can solve problems such as difficult to use, and achieve high reliability, consistency and probability of definite results high effect

Inactive Publication Date: 2011-06-15
UNIV DE CONCEPCION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, none of these patents are related to more than 2 virulence genes, at most one identified sequence, or they are difficult techniques to apply

Method used

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  • Tjn molecular kit for diagnosing virulent strains of helicobacter pylori
  • Tjn molecular kit for diagnosing virulent strains of helicobacter pylori
  • Tjn molecular kit for diagnosing virulent strains of helicobacter pylori

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0091] Example 1: The kit is applied to gastric biopsy samples

[0092] This is not a limiting example of the technology, but only a specific application of the kit developed.

[0093] sample:

[0094] Fifty-six biopsies were analyzed from the gastric antrum and gastric body of 26 patients who had seen a doctor for gastric and duodenal problems. Samples of H. pylori strains ATCC 43504, ATCC 25695 and 96.978 were used as positive controls and samples of human genomic DNA were used as negative controls.

[0095] 1- First DNA extraction phase and secondary phases:

[0096] DNA was extracted using the reactions described below (available in the proposed kit): lysis buffer (TrisHCl 10 mM, EDTA 1 mM, SDS 10%); proteinase K (20 mg / mL); CTAB / NaCl solution; chloroform:iso Pentanol (24:1); Benzene:chloroform:isoamyl alcohol=25:24:1; ethanol and TE buffer. Fifty biopsies analyzed were included in the assay; other samples were used as controls and DNA was extracted from pure isolates ...

Embodiment 2

[0102] Embodiment 2: the comparison of the present invention and commercial kit

[0103] Our technology was compared with an alternative commercial kit MPCR H. pylori kit Cat N°. MP-700081. The latter showed significant differences in terms of results, as described below:

[0104] 1) The latter does not contain the reactions or steps required to extract DNA from gastric biopsy samples and / or H. pylori cultures.

[0105] 2) The latter does not provide the reactants Taq polymerase and dNTP, which means that kit users must prepare these substances in advance. Furthermore, it includes reactants in the form of sterile liquids, which are prone to loss of activity during temperature changes (they must be stored at -20°C and shipped at -20°C). Our invention includes a choice of two formats: sterile lyophilized beads (completely stable for storage and transport at ambient temperature) or sterile liquid.

[0106] 3) MPCR Helicobacter pylori kit Cat N°.MP-700081 detects gene ureA, acc...

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Abstract

A kit in the form of a product and a method able to detect simultaneously four genes of Helicobacter pylori (rDNAl6S Hpy), i.e. one identification gene and three virulence genes (cagA, vacAml, dupA), is disclosed. Moreover, the kit envisages the association of primers which determine the quality of extraction of the DNA (Eub gene).

Description

Background technique [0001] Infections caused by Helicobacter pylori are a major public health problem because their control is very expensive. This situation has led to a need for tools to detect risk factors associated with this condition. However, there are currently no commercially available microbial assays to detect virulent strains. [0002] Since there are currently no reliable laboratory tools for the detection of H. pylori, eradication therapy for this bacterium is based only on qualitative methods, and there are no treatments or treatments to measure the virulence of H. pylori. [0003] Existing methods only indicate the presence or absence of microorganisms, but do not reveal the nature or identity of the infecting strain. Another important aspect is that only very sporadic endoscopic studies have been performed in children, so there are very few data on infections in this age group. [0004] According to reports worldwide, eradication therapy fails in 20% to 80...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12P19/34
CPCC12Q1/689C12Q2600/16
Inventor A·加西亚坎西诺C·冈萨雷斯科雷亚N·特拉巴尔费尔南德斯
Owner UNIV DE CONCEPCION
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