Specificity detection primers of Aids related intestinal tract Megamonas funiformis translocation and application

A technology for detecting primers and AIDS, which is applied in the field of molecular biology, can solve the problems of unfavorable early diagnosis and treatment, unreliable results, delayed diagnosis and other problems, and achieve the effect of good practicability, reliable results and small sample size

Inactive Publication Date: 2016-09-21
FIRST AFFILIATED HOSPITAL OF KUNMING MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the diagnosis of intestinal flora translocation is mainly based on the basic disease and clinical manifestations of the patient plus laboratory diagnosis. The commonly used method is the detection of endotoxin in the blood of the patient. Although this method is fast, it cannot be specific. The type of bacteria infect

Method used

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  • Specificity detection primers of Aids related intestinal tract Megamonas funiformis translocation and application
  • Specificity detection primers of Aids related intestinal tract Megamonas funiformis translocation and application

Examples

Experimental program
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Embodiment 1

[0027] Example 1: Primer Design

[0028] Design based on 16S rRNA hypervariable region Megamonas funiformis A pair of specific primers, the target band of PCR amplification is 517bp, and the primer sequence is:

[0029] .

Embodiment 2

[0030] Example 2: Specific detection primers for detection of intestinal Prevotella translocation in AIDS patients

[0031] The test group of this embodiment is the peripheral blood DNA of 19 cases of AIDS patients; the peripheral blood DNA of the 19 cases of AIDS patients is sequenced by the second-generation high-throughput 16S rRNA sequencing method at the same time as a comparison; the peripheral blood DNA of 20 cases of healthy people is used as For the control group, replace the template with water as the blank control; Megamonas funiformis DNA was used as a positive control.

[0032] 1. Preparation of DNA template

[0033] Blood DNA was extracted using a commercially available blood DNA extraction kit and operated according to the instructions of the kit.

[0034] 2. PCR reaction and gel electrophoresis

[0035] Reaction system: Gold Mix (TSE101) kit, the reaction system is 50 μL system, less than 50 μL is made up with double distilled water, the details are as foll...

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Abstract

The invention discloses specificity detection primers of Aids related intestinal tract Megamonas funiformis translocation and application. The sequence of an upstream primer is Meg-F5'-AGTTCCTTCTCTTCGGAGAAC-3', and the sequence of a downstream primer is Meg-R5'-TAAGGAGGTGATCCAGCC-3'; the specificity detection primers have the advantages that the detection is accurate, simple, convenient and rapid, the sensitivity is high, and the specificity is high and have excellent detectability and a wide application prospect; the technology can be widely applied to early stage monitoring before symptom appearance of Aids patient intestinal tract Megamonas funiformis flora translocation, and a scientific basis is provided for formulating the treatment strategy.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a specific detection primer set and application for detecting the translocation of the intestinal megamonas fusiformis of AIDS patients. Background technique [0002] AIDS, or Acquired Immune Deficiency Syndrome, has been on the rise internationally. As far as the country is concerned, as of the end of October 2015, a total of 575,000 HIV-infected people and patients have been reported nationwide. , 177,000 people died. In recent years, more and more experts and scholars have shifted the focus of AIDS research to the intestinal tract. It is generally believed that the intestinal tract is the target organ for HIV to attack and replicate, and the intestinal flora disorder and intestinal barrier damage caused by the same time make the intestinal tract The translocation of flora induces a series of complications such as infection and chronic immune system activation, thus accelerati...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04C12N15/11
CPCC12Q1/689
Inventor 王昆华谢振荣徐玉任莉沈宗文王腊梅王华伟
Owner FIRST AFFILIATED HOSPITAL OF KUNMING MEDICAL UNIV
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