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Saliva VOC marker of helicobacter pylori and application of saliva VOC marker in preparation of diagnostic reagent

A technology of Helicobacter pylori and markers, which is applied in the direction of instruments, measuring devices, scientific instruments, etc., can solve the problem that VOC marker components are easily adsorbed on the inner wall of the gas collection bag, the storage time is difficult, the breath samples are not easy to store and transport, and the occurrence of gastric cancer can be reduced Efficiency and other issues, to achieve the effect of easy storage and transportation, rapid collection, and little matrix effect interference

Pending Publication Date: 2022-07-29
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] However, in the detection technology of Helicobacter pylori disclosed in the prior art, such as in the patent CN113866307A, the method of diagnosis based on the VOC markers in the breath sample, the breath sample is not easy to store and transport, and the VOC markers in the breath The components are easily adsorbed on the inner wall of the gas sampling bag or changed due to long storage time; patent CN113984948A is to compare and screen VOC markers for Helicobacter pylori strains and gastric mucosal epithelial cells, which requires sampling of gastric mucosal tissue, strains and cells Culture, the steps are cumbersome, professional and the sampling process will bring discomfort to the subjects; while the patent CN110045035B discloses the application of a VOC marker in saliva in the diagnosis of gastric cancer, Helicobacter pylori infection is considered to cause A possible cause of gastric cancer, so timely diagnosis at the stage of Helicobacter pylori infection is expected to effectively reduce the incidence of gastric cancer

Method used

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  • Saliva VOC marker of helicobacter pylori and application of saliva VOC marker in preparation of diagnostic reagent
  • Saliva VOC marker of helicobacter pylori and application of saliva VOC marker in preparation of diagnostic reagent

Examples

Experimental program
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Effect test

Embodiment 1

[0032] Collection of saliva samples

[0033]None of the 46 volunteers who provided saliva (17 cancer patients and 29 normal subjects) had other digestive or metabolic diseases. The saliva of each subject was analyzed using a total of 46 saliva samples. Forty-six volunteers were confirmed as positive and negative groups of Helicobacter pylori infection through C14 breath test and gastroscope to take gastric mucosa specimens for bacterial culture in advance for double verification. Before sample collection, each volunteer should maintain a natural and peaceful mood within 48 hours, and refrain from smoking and drinking within 24 hours. Information such as age, gender, detection value of Helicobacter pylori, history of smoking and drinking, current drug use, and diet in the past three days were recorded for each volunteer. All volunteers were forbidden to eat any food for 1 hour before sample collection to keep their mouth clean. Before sampling, gargle with pure water, and st...

Embodiment 2

[0035] Pretreatment and detection of saliva samples

[0036] Put the obtained saliva sample into a centrifuge tube, centrifuge at 12,000 rpm for 15 minutes, remove food residues and other magazines, accurately control the injection volume to 1 ml, and heat the sample for 30 minutes to 65 degrees to ensure that the vapor reaches chemical equilibrium . The headspace sampling device was connected to the solid-phase microextraction device, and a 75-micron CAR / PDMS-coated solid-phase microextraction head was used to pre-concentrate the markers in the sample. The VOCs in the samples were pre-concentrated, and the concentration time was respectively 30 minutes. GC-MS analysis: The extraction head was placed in the GC-MS injection port, the high temperature was 280 degrees, the desorption was instantaneously desorbed for 5 minutes, and the sample was injected in a splitless manner. After 5 minutes, open the diverter valve. Chromatographic separation was performed on a DB-WAX column...

Embodiment 3

[0038] Screening of Helicobacter pylori infection markers in saliva samples

[0039] The detected substances were preliminarily identified using the NIST14 library of mass spectrometry, and substances with a similarity of more than 90% were quantified using the percentage of peak area content. Chromatograms, content comparison charts and corresponding specific VOC markers of saliva samples of Helicobacter pylori infection positive and negative groups such as figure 1 , figure 2 and shown in Table 1.

[0040] Table 1 VOC markers associated with H. pylori infection

[0041] Peak number Chemical Name 1 acetone 2 2-Methyl-2-propanol 3 Ethanol 4 2-Pentanone 5 1-Propanol 6 2-Methyl-1-propanol 7 2-Pentanol 8 1-Butanol 9 2-Methyl-1-butanol 10 acetoin 11 Pyrrole 12 p-Tolualdehyde

[0042] The application of diagnosing Helicobacter pylori infection by VOC markers in saliva, when the peak area per...

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Abstract

The invention discloses a VOC (volatile organic compound) marker for helicobacter pylori in saliva, which is characterized in that the VOC marker is a combination of acetone, 2-methyl-2-propanol, ethanol, 2-pentanone, 1-propanol, 2-methyl-1-propanol, 2-pentanol, 1-butanol, 2-methyl-1-butanol, acetoin, pyrrole and p-tolualdehyde. The diagnostic reagent is used for non-label diagnosis of helicobacter pylori infection by detecting the content of the helicobacter pylori related VOC marker in saliva of a testee, and the multiple volatile organic compounds in saliva are used as markers for diagnosis of helicobacter pylori infection. The application prospect in screening and diagnosis of the helicobacter pylori is wide.

Description

technical field [0001] The invention relates to the technical field of detection of Helicobacter pylori, in particular to a VOC marker in saliva and its application in the preparation of diagnostic reagents. Background technique [0002] Helicobacter pylori is a pathogenic bacteria in the digestive system. It is a microaerophilic gram-negative bacterium that can survive in the extremely low pH acidic environment in the stomach, causing about 50% of the world's population to infect. Helicobacter pylori has been confirmed to be closely related to gastrointestinal diseases such as chronic active gastritis, gastric ulcer and even gastric cancer, gastric mucosa-associated lymphoma, and is listed as the first carcinogen. [0003] At present, the diagnosis methods for Helicobacter pylori infection mainly include carbon 13 / 14-labeled urea breath test, stool antigen test, serological antibody test, urine antibody test, and invasive tests such as endoscopy, histological test, Immunoh...

Claims

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Application Information

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IPC IPC(8): G01N30/88
CPCG01N30/88G01N2030/8813
Inventor 崔大祥薛翠丽金涵徐余丽张禹娜张正虎
Owner SHANGHAI JIAO TONG UNIV
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