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

Application of product for detecting ddcfDNA in preparing product for detecting transplanted kidney injuries caused by pulmonary infection and evaluating prognosis effect

A technology for lung infection and kidney injury, applied in the field of medical detection, can solve the problems of difficulty in evaluating the effect of lung infection on transplanted kidney injury, inability to effectively detect transplanted kidney injury and prognosis, etc. Painful effects of patients

Pending Publication Date: 2019-11-15
上海奥根诊断技术有限公司
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Therefore, the problem to be solved by the present invention is to overcome the inability to effectively detect the injury and prognosis of the transplanted kidney caused by pulmonary infection in existing clinics, and it is difficult to evaluate the damage of pulmonary infection to the transplanted kidney and the effect after treatment of pulmonary infection, and then provide a A product for detecting ddcfDNA in the preparation of a product for detecting lung infection on transplanted kidney injury and prognosis, a product for detecting lung infection on transplanted kidney injury and prognosis, and detecting the effect of lung infection on transplanted kidney injury and prognosis method of prognosis

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 product for detecting ddcfDNA in preparing product for detecting transplanted kidney injuries caused by pulmonary infection and evaluating prognosis effect
  • Application of product for detecting ddcfDNA in preparing product for detecting transplanted kidney injuries caused by pulmonary infection and evaluating prognosis effect
  • Application of product for detecting ddcfDNA in preparing product for detecting transplanted kidney injuries caused by pulmonary infection and evaluating prognosis effect

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Example 1 SNP site screening

[0070] Select the dbSNP (data base SNP, https: / / www.ncbi.nlm.nih.gov / snp / ) database and the Chinese population database (saved in Shanghai Aogen Diagnostics Technology Co., Ltd.), the Chinese population database is based on 3000 cases of Chinese The catalog of genetic polymorphism sites constructed from the whole genome sample information, such as figure 1 As shown, there are some differential frequency information between the Chinese population database and the Hapmap database (http: / / www.hapmap.org), which represent the differential genetic loci in the Chinese population, enabling the Chinese population database to more accurately present the Chinese population genetic polymorphism information.

[0071] Sites with a minor allele frequency (MAF) close to 0.5 were screened from the above two databases, and 5754 target SNP sites shown in Table 1 were obtained.

[0072] The SNP sites of the drug metabolism genes, such as CYP3A4, CYP3A5 and...

Embodiment 2

[0095] Example 2 Extraction of Genomic DNA and Plasma cfDNA

[0096] 1. Sample collection and separation of blood cells and plasma

[0097] (1) Sample collection: The blood drawn from the donor is divided into two parts, about 8ml each, stored and transported in streck tubes;

[0098] (2) Plasma separation: centrifuge a fresh whole blood sample at 1900g at 4°C for 10min, separate the supernatant into a new centrifuge tube, continue to centrifuge the supernatant at 16000g at 4°C for 10min, and centrifuge the supernatant Transfer the solution to a new centrifuge tube to obtain separated plasma, which can be stored at -20°C;

[0099] (3) Blood cell separation: the lower sediment after centrifugation of another fresh whole blood sample is blood cells.

[0100] 2. Use (Laifeng Genomic DNA Extraction Kit, DK601-02) to extract the separated blood cells to obtain genomic DNA; use (QIAamp Circulating Nucleic Acid Kit, 55114) to extract the separated plasma to obtain plasma cfDNA, the...

Embodiment 3

[0107] Example 3 library construction

[0108] 1. Take 1 μl of the plasma cfDNA obtained in Example 2 for QuantiFluor TM - Quantification by ST (Promega), take another 1 μl and use Agilent 2100 to check the quality.

[0109] 2. Using plasma cfDNA and blood cell genomic DNA obtained in Example 2 as samples, use VAHTSUniversal DNA Library Prep Kit for Illumina V3 to prepare plasma cfDNA and genomic DNA libraries. The specific steps are as follows:

[0110] (1) End filling

[0111] a. Add 7.5 μl End Repair mix to the marked centrifuge tube;

[0112] b. Take 25 μl of the sample quantified in step 2, add the end-filling mixture shown in a to obtain a sample reaction solution with a total volume of 32.5 μl, and mix with a pipette;

[0113] c. Run the following program on the PCR machine:

[0114] Keep at 20°C for 15 minutes,

[0115] Keep at 65°C for 15 minutes,

[0116] Stand at 4°C.

[0117] (2) Add single-molecule marker adapter

[0118] a. According to the number of samp...

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

No PUM Login to View More

Abstract

The invention provides application of a product for detecting ddcfDNA in preparing a product for detecting transplanted kidney injuries caused by pulmonary infection and evaluating a prognosis effect,the product for detecting the transplanted kidney injuries caused by pulmonary infection and evaluating the prognosis effect, and a method for detecting the transplanted kidney injuries caused by pulmonary infection and evaluating the prognosis effect. The product for detecting the ddcfDNA can detect the level of the donor free cfDNA (ddcfDNA) in plasma of a kidney transplant recipient, accordingto the level of the donor free cfDNA (ddcfDNA), the transplanted kidney function injuries caused by pulmonary infection are diagnosed, the prognosis effect is evaluated, the early diagnosis of the transplanted kidney injures caused by pulmonary infection and the prognosis effect for the organ transplant recipient after an operation can be achieved, the product for detecting the ddcfDNA has the advantages of rapidity, accuracy and high sensitivity, the detection method is non-invasive, and the pain of the patient is greatly reduced.

Description

technical field [0001] The invention relates to the technical field of medical detection, in particular to the use of a product for detecting ddcfDNA in the preparation of a product for detecting transplanted kidney injury and prognosis caused by lung infection, for detecting the effect of lung infection on transplanted kidney injury and prognosis products, and methods for detecting the impact of pulmonary infection on transplanted kidney injury and prognosis. Background technique [0002] Infection is one of the most common complications after organ transplantation. Although with the development and use of new immunosuppressive drugs, which can reduce the incidence of rejection and prevent and treat infectious complications, the survival rate of organ transplant recipients has increased significantly in the past few decades. However, infection is still an important cause of organ transplant failure. [0003] Immunosuppressants are used in patients after renal transplantat...

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
IPC IPC(8): C12Q1/6883C40B50/06C12Q1/6869
CPCC12Q1/6883C40B50/06C12Q1/6869C12Q2600/118C12Q2600/156C12Q2535/122
Inventor 蒋廷亚刘海涛葛军刘枫王啸晨黎小锋
Owner 上海奥根诊断技术有限公司
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

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