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Primer pair, kit and method for detecting whether people smoke or not based on saliva microorganisms

A microbial detection and microbial technology, applied in the field of bioinformatics, can solve problems such as concealing the truth, and achieve the effect of overcoming difficult cultivation, easy acquisition and preservation

Active Publication Date: 2020-07-10
CHINA TOBACCO YUNNAN IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] .Currently, the information about whether the subject smokes can only be obtained by asking, but whether it is the subject himself or others has the possibility of concealing the truth

Method used

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  • Primer pair, kit and method for detecting whether people smoke or not based on saliva microorganisms
  • Primer pair, kit and method for detecting whether people smoke or not based on saliva microorganisms
  • Primer pair, kit and method for detecting whether people smoke or not based on saliva microorganisms

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] A primer pair for detecting whether people smoke based on saliva microorganisms, including an upstream primer and a downstream primer;

[0042] The upstream primer is sequentially composed of a tag sequence and an upstream primer sequence;

[0043] The downstream primer consists of a tag sequence and a downstream primer sequence in turn;

[0044] The sequence of the upstream primer is shown in SEQ ID NO: 1; the sequence of the downstream primer is shown in SEQ ID NO: 2; the tag sequence is composed of random bases.

[0045] A kit for detecting whether a crowd smokes based on saliva microorganisms, the kit includes the above-mentioned primer pair, PCR reagent and negative control.

Embodiment 2

[0047] A primer pair for detecting whether people smoke based on saliva microorganisms, including an upstream primer and a downstream primer;

[0048] The upstream primer is sequentially composed of a tag sequence and an upstream primer sequence;

[0049] The downstream primer consists of a tag sequence and a downstream primer sequence in turn;

[0050] The sequence of the upstream primer is shown in SEQ ID NO: 1; the sequence of the downstream primer is shown in SEQ ID NO: 2; the tag sequence is randomly composed of 5 bases.

[0051] A kit for detecting whether a crowd smokes based on saliva microorganisms, the kit includes the above-mentioned primer pair, PCR reagent and negative control.

[0052] The PCR reagent is Taq DNA polymerase (2X). Preferred is Invitrogen Platinum Taq DNA Polymerase (2X).

[0053] The negative control is the oral saliva microbiome of non-smokers.

Embodiment 3

[0055] A method for detecting whether a person smokes based on saliva microorganisms, using the primer pair of Example 1 or the kit of Example 1, comprising the steps of:

[0056] Step (1), sample collection: use sterilized EP tubes to collect 3-5mL saliva samples from the person to be tested, immediately put them into liquid nitrogen for quick freezing for 5 minutes, and store them at -80°C;

[0057] Step (2), extracting total DNA: using the CTAB method to extract the total DNA in saliva;

[0058]Step (3), amplification of the target fragment: PCR amplification is performed using the total DNA as a template, the PCR product is sequenced after recovery and purification, and OTUs clustering and species classification analysis are performed;

[0059] Step (4), judging whether the person is a smoker or not: when the following three conditions are met at the same time, the person to be detected is a smoker;

[0060] ① The microbial diversity of the sample to be tested is signific...

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Abstract

The invention relates to a primer pair, a kit and a method for detecting whether people smoke or not based on saliva microorganisms, and belongs to the technical field of bioinformatics. The kit comprises a primer pair, a PCR reagent and a negative control; the primer pair comprises an upstream primer and a downstream primer; the upstream primer consists of a tag sequence and an upstream primer sequence in sequence; the downstream primer consists of a tag sequence and a downstream primer sequence in sequence; the sequence of the upstream primer is shown as SEQ ID NO: 1; and the sequence of thedownstream primer is shown as SEQ ID NO: 2. The method comprises the steps of firstly collecting a saliva sample of a person to be detected, then extracting total DNA in saliva, then carrying out PCRamplification, carrying out sequencing after a PCR product is recovered and purified, carrying out clustering and species classification analysis of OTUs, and then carrying out judgment according toan analysis result. The method can effectively predict whether the crowd has a long-term smoking habit, and the detection process is noninvasive, low in cost, rapid, convenient, high in safety and easy to popularize and apply.

Description

technical field [0001] The invention belongs to the technical field of bioinformatics, and in particular relates to a primer pair, a kit and a method for detecting whether a crowd smokes based on saliva microorganisms. Background technique [0002] There are more than 600 kinds of bacteria in the human oral cavity, which are collectively referred to as the oral microbiome. The human oral microbiome is the second most diverse and community-rich microbiome in the human body. These bacteria have many functions, many of which are important in maintaining oral health. Abnormal oral microbiome may lead to periodontitis, dental caries and other oral diseases. There is also increasing evidence that dysbiosis of the oral microbiome also plays an important role in the development of diseases of the lung, gastrointestinal tract, and cardiovascular system. Smoking and alcohol consumption are important influencers of the oral microbiome, affecting microbial diversity and their underlyi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/6806C12Q1/04C12N15/11C12R1/01
CPCC12Q1/689C12Q1/6806C12Q2531/113C12Q2535/122Y02A50/30
Inventor 高茜米其利杨继管莹曾婉俐向海英杨光宇李雪梅
Owner CHINA TOBACCO YUNNAN IND
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