Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Application of mir-671-5p reagent for detection of human serum exosomes and osteoporosis detection kit

A technology for osteoporosis and exosomes, applied in medical testing and biological fields, can solve the problems of inability to accurately quantify the molecular copy number, the specificity of the dye method is not as good as the probe method, and the lower limit of detection and sensitivity are unknown. The effect of simplifying the experimental steps, improving specificity and sensitivity, and improving efficiency

Active Publication Date: 2022-05-31
THE SECOND XIANGYA HOSPITAL OF CENT SOUTH UNIV
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Extract exosomes from serum and use fluorescent PCR to detect miR-671-5p. Currently, there are dye method and probe method. The dye method is not as specific as the probe method. At present, these two methods are relatively quantified by the internal reference Ct value The method cannot accurately quantify the specific molecular copy number, and the detection limit and sensitivity are unknown
Moreover, the current existing methods are all single-channel single-color detection, and microRNA and internal reference each need a separate tube for PCR, which results in low detection efficiency and high cost.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Application of mir-671-5p reagent for detection of human serum exosomes and osteoporosis detection kit
  • Application of mir-671-5p reagent for detection of human serum exosomes and osteoporosis detection kit
  • Application of mir-671-5p reagent for detection of human serum exosomes and osteoporosis detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] (5) Add 500 μL of washing solution 1 to the column, and vacuum suction for 1 minute. Add 500 μL of Wash 2 to the column and vacuum

Embodiment 2

(1) Take 3× reverse transcription mixed solution 5 μL, 0.5 μL of reverse transcription primer mixed solution of 1 μmol / L and add 0.2 mL RNA-free

Enzymes in PCR thin-walled tubes.

[0077] (2) Take 9.5 μL of the extracted microRNA, add it to the above-mentioned mixed solution, and mix by pipetting.

[0078] (3) Put into a PCR machine for reverse transcription, and the conditions are: 42°C for 30 minutes and 70°C for 5 minutes. Store the obtained cDNA in an ice box

Embodiment 3

[0081] (2) Add 15 μL of reverse transcribed cDNA to each tube and mix well.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
diameteraaaaaaaaaa
Login to View More

Abstract

The present invention provides an application of detecting miR-671-5p reagent in human serum exosomes and an osteoporosis detection kit, which includes reverse transcription primers of miR-671-5p, fluorescent probes, and PCR primers, and can also be It further includes internal reference reverse transcription primers, fluorescent probes and PCR primers, and the kit further includes miR-671-5p reference and internal reference. The kit uses double fluorescent quantitative PCR to detect the absolute concentration of exosomal miR-671-5p from a small amount (200 μL) of serum or plasma. The detection method is specific, efficient and sensitive. The kit can be used for the prediction and evaluation of osteoporosis in the elderly, and has important clinical significance.

Description

Application of miR-671-5p reagent for detecting human serum exosomes and osteoporosis detection Reagent test kit technical field The present invention relates to biology, medical detection field, particularly detect miR-671-5p reagent in human serum exosomes application and osteoporosis detection kit. Background technique The prevalence rate of the elderly over 60 years old in my country is 36%, and there may be no obvious symptoms in the initial stage, which is called "silent disease", and it is difficult to For early detection and diagnosis, currently only bone density testing can be used for screening, prevention is better than treatment, and there is no better detection method. Exosome (exosome) is a kind of lipid bilayer structure, nano-scale vesicles secreted from the inside to the outside of the cell. The diameter is 30-150nm, containing specific proteins, lipids, nucleic acids and other substances, and various components it contains mediate the relationship...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883C12Q1/686C12N15/113
CPCC12Q1/6883C12Q1/686C12N15/113C12N2310/141C12Q2600/158C12Q2600/178C12Q2600/166C12Q2563/107C12Q2537/143
Inventor 张旻宋德业蒋永芳周宁肖新强
Owner THE SECOND XIANGYA HOSPITAL OF CENT SOUTH UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products