Plasma miRNA (micro ribonucleic acid) marker associated with common type congenital intestinal canal total colonic aganglionosis and application thereof
A technology of ganglion cells and markers, applied in the fields of genetic engineering and clinical medicine, can solve problems such as lack of corresponding attention
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0086] Embodiment 1 Research Object Selection and Grouping Basis
[0087] From July 2009 to September 2011, the inventor collected children with common congenital intestinal aganglionosis and normal non-congenital intestinal aganglionosis from the Children's Hospital Affiliated to Nanjing Medical University and other hospitals that met the requirements. Blood and tissue samples (for clinical diagnostic criteria see figure 1 , figure 2), from which 100 healthy controls (average age: 89.45±9.1 days) and 100 patients with intestinal common congenital intestinal aganglionosis (average age: 92.3±7.01 days) were selected to meet the requirements ) as the experimental object of Real-time PCR detection of miRNA expression. Specific sample classification criteria are as follows:
[0088] Group A: healthy control group (n=100, 20 people for microarray screening, 40 people for first-stage verification, 40 people for independent crowd verification):
[0089] 1. Age between 0 and 6 mo...
Embodiment 2
[0100] Embodiment 2 research object pathological diagnosis type
[0101] Anorectal manometry, imaging examination, and rectal wall histological examination were used to diagnose intestinal aganglionosis and preliminarily clarify its type for children with delayed meconium discharge, repeated defecation difficulties, abdominal distension, and low intestinal obstruction in the past. Anorectal manometry mainly shows the absence of internal anal sphincter relaxation reflex; the rhythmic contraction of anal canal is significantly reduced; the resting pressure of internal rectal sphincter is higher than normal. Imaging examination showed that: 1. There was an obvious transition zone between the lesion segment and the dilation segment, showing a "cone" shape; 2. The lesion segment extended upward from the anus to the distal end of the sigmoid colon; 3. The innervation of the lesion segment was abnormal Therefore, it can be seen that there are irregular contractions; 4. The retention ...
Embodiment 3
[0102] Example 3 Taqman miRNA array screening
[0103] Preparation of cDNA samples: a) Take 100 μl plasma; b) Add 900 μl Trizol, shake and mix well, centrifuge at 12,000 rpm at 4°C for 15 minutes, discard the lower layer of waste liquid; c) Add 1.5 times the volume of supernatant absolute ethanol, shake and mix, transfer To the spin column, centrifuge at 12000 rpm for 15 seconds, discard the lower waste liquid; d) add 700 μl RWT buffer to the spin column, centrifuge at 10000 rpm for 15 seconds, discard the lower waste liquid. e) Add 500 μl of RPE buffer to the spin column, centrifuge at 10,000 rpm for 15 seconds, and discard the lower layer. f) Repeat e. g) Add the spin column to a new 2ml tube and centrifuge at 10,000rpm for 1 minute to remove the RPE buffer. h) Add 50 μl of DEPC-treated water to the column and collect RNA by centrifugation at 12000 rpm. i) Then cDNA is obtained by RNA reverse transcription reaction. The reverse transcription reaction system included 4 μl...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com