Application of intestinal microorganism metabolite in preparation of autism diagnosis kit

A technology of intestinal microorganisms and diagnostic kits, applied in biological testing, disease diagnosis, biomaterial analysis, etc., can solve the problems of no molecular biology diagnostic standards, low diagnostic accuracy, and limited development of treatment methods, and achieve important medical applications Prospect and economic value, high diagnostic accuracy, effect of high accuracy

Active Publication Date: 2020-11-24
THE SEVENTH AFFILIATED HOSPITAL SUN YAT SEN UNIV SHENZHEN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

On the other hand, so far, the diagnostic process of autism often relies on scales for evaluation, and there is no diagnostic standard in molecular biology
This type of diagnostic method is easily interfered by subjective experience, resulting in low diagnostic accuracy and limiting the development of related treatments.

Method used

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  • Application of intestinal microorganism metabolite in preparation of autism diagnosis kit
  • Application of intestinal microorganism metabolite in preparation of autism diagnosis kit
  • Application of intestinal microorganism metabolite in preparation of autism diagnosis kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Determination of acetic acid content in the stool sample of embodiment 1

[0024] In this example, stool samples from healthy people and patients with autism were detected, and then correlation analysis was performed to determine the correlation between the level of acetic acid, an intestinal microbial metabolite in stool samples, and the onset and severity of symptoms of autism. Specific steps are as follows:

[0025] (1) Sample source

[0026] Autism group: from Shanxi Zhou Autism Rehabilitation Research Institute, age distribution 0-14 years old. Inclusion criteria: ①According to the latest international diagnostic criteria, that is, "Diagnostic and Statistical Manual of Mental Disorders, 5th Edition, DSM5", a total of 49 patients diagnosed with autism, of which Mild in 41 cases, severe in 8 cases; ②Gender is not limited. Exclusion criteria: ① suffering from other psychiatric diseases (such as schizophrenia) and other neurodevelopmental disorders; ② suffering from...

Embodiment 2

[0040] Determination of content of tyramine, histamine, γ-aminobutyric acid, dihydroxyphenylalanine and norepinephrine in the stool sample in embodiment 2

[0041] Continue to analyze other intestinal microbial metabolites on the collected stool samples. The specific steps are as follows:

[0042] (1) Preparation of the second sample (the second sample and the first sample in Step (3) of Example 1 are two samples derived from the same stool sample)

[0043] Weigh an appropriate amount for each portion, add 1% FA 200 μL pre-cooled ultrapure water, perform MP homogenization, then add 1% FA 800 μL methanol / acetonitrile (1:1, v / v), vortex mix, low-temperature ultrasound for 30 minutes, - Incubate at 20°C for 1h to precipitate protein, centrifuge at 14000rcf at 4°C for 20min, take the supernatant and dry it in vacuum. Add 1% FA in 100 μL ACN / water (1:1, v / v) for reconstitution during mass spectrometry detection, centrifuge at 14000 rcf at 4°C for 15 min, and take the supernatant f...

Embodiment 3

[0050] Example 3 Correlation Analysis of the Content of Gut Microbial Metabolites in Stool Samples and Autism

[0051] Utilize PLS-DA analysis (Partial Least Squares Discriminant Analysis) to measure the content of intestinal microbial metabolites in the stool samples of the autism group and control group measured in Example 1 and Example 2.

[0052] The result is as figure 1 As shown, the two groups of samples (autism group and control group) were divided into two circles, indicating that the overall expression patterns of metabolites in the two groups were different, so the concentration of microbial intestinal metabolites can significantly distinguish healthy subjects. and autistic patients (11.28% of the explained variance).

[0053] Such as figure 2 Shown is the variable importance for the projection (Variable Importance for the Projection, VIP) in the PLS-DA analysis. VIP is the variable weight value of the PLS-DA model variable, which can be used to measure the impa...

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Abstract

The invention discloses an autism diagnosis kit, the autism diagnosis kit comprises a reagent for detecting the intestinal microbial metabolite concentration, wherein the intestinal microbial metabolite comprises at least one of acetic acid, tyramine, histamine, gamma-aminobutyric acid, dihydroxyphenylalanine and norepinephrine. The acetic acid content and the score of the severity degree of the autism are in a negative correlation relationship; the contents of tyramine, histamine, gamma-aminobutyric acid, dihydroxyphenylalanine and norepinephrine are respectively in positive correlation withthe severity of autism, and the autism is diagnosed through the intestinal microbial metabolite level of an excrement sample, so that the kit has relatively high accuracy. Therefore, the intestinal microorganism metabolite combination level of a subject can be used as an index for screening and diagnosing autism under the conditions of scientific research and clinical application.

Description

technical field [0001] The invention relates to the field of autism diagnosis, in particular to the use of intestinal microbial metabolites in the preparation of autism diagnosis kits. Background technique [0002] Autism spectrum disorder (autism spectrum disorder, ASD), referred to as autism, also known as autism spectrum disorder or autism, is a generalized neurodevelopmental disorder, mainly manifested as social impairment, language impairment and repetitive and Stereotyped behavior. According to the survey results of "Report on the Development of China's Autism Education and Rehabilitation Industry" (2015 Edition), there are about 10 million autistic patients in China (according to the incidence rate of 1%), including about 2 million children (0-14 years old). age). Autism has brought a heavy burden to the social public health system and patients' families. [0003] Autism is a neurocognitive disorder with complex etiology, and its pathogenesis is influenced by genet...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/72G01N33/68G01N33/94
CPCG01N30/02G01N30/72G01N33/6893G01N33/9433G01N2800/28
Inventor 李宁宁王迪龙潘逸航
Owner THE SEVENTH AFFILIATED HOSPITAL SUN YAT SEN UNIV SHENZHEN
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