Gene detection composition and its application

A composition and gene technology, applied in the CCDC72 and ERC1 gene composition, the detection of postmenopausal osteoporosis and the clinical application of Liuwei Dihuang Pill, TMED2, the field of gene detection composition, can solve the unproven correlation, etc. problems, to achieve the effect of simplifying the quantitative detection process, high sensitivity, and clear results

Active Publication Date: 2020-04-10
GUAN BOJIAN BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] There are not many studies on the ERC1 gene. In 2016, an article on the screening of genes related to congenital diaphragmatic hernia showed that the ERC1 gene may be related to congenital diaphragmatic hernia, but the correlation has not been confirmed (screening of genes related to congenital diaphragmatic hernia, Shandong Medicine, 2016 Volume 56, Issue 19)

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  • Gene detection composition and its application
  • Gene detection composition and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1 High-throughput sequencing and analysis

[0050] The peripheral blood samples of 3 postmenopausal osteoporosis patients before and after taking Liuwei Dihuang Pills and 3 healthy controls were collected for RNA extraction. After RNA extraction, agarose gel electrophoresis was performed. Whether the quality of the RNA sample is qualified or not, whether it can be used for further transcriptome analysis. Then, the extraction of RNA samples was detected by NanoDrop1000 spectrophotometer, and the sample requirements for RNA-seq sequencing: OD260 / OD280 was 1.8-2.2.

[0051] The sequencing platform is the HiSeq 2500 high-throughput sequencing platform of Illumina, which performs high-throughput transcriptome deep sequencing. After sequencing, we use Fast-QC software to evaluate the quality of the sequencing data as a whole, including the quality value distribution of bases, quality value position distribution, GC content, PCR duplication content, kmer frequency, et...

Embodiment 2

[0052] Example 2 Peripheral blood TMED2, CCDC72 and ERC1 gene expression in postmenopausal osteoporosis patients before and after taking Liuwei Dihuang Pills

[0053]1. Materials and methods

[0054] 1. Materials

[0055] The consent of 22 patients with postmenopausal osteoporosis was collected, and the peripheral blood was collected at the initial stage of testing. After taking Liuwei Dihuang Pills for half a year, the peripheral blood was collected again, and the samples were grouped and numbered.

[0056] 2. Method

[0057] 2.1 Extraction of total RNA from peripheral blood of postmenopausal osteoporosis and healthy people

[0058] use Reagent was used to extract RNA from samples, and the experimental operation was performed according to the product instructions.

[0059] Judgment criteria for RNA quality: the OD260 / OD280 value of the RNA sample is between 1.8 and 2.2; the electrophoretic pattern of total RNA has clear 28S and 18S bands; difference.

[0060] 2.2 Synth...

Embodiment 3

[0083] Example 3 TMED2, CCDC72 and ERC1 gene expression in peripheral blood of postmenopausal osteoporosis patients and healthy people

[0084] 1. Materials

[0085] The peripheral blood of 32 postmenopausal osteoporosis patients and 29 healthy people were collected, grouped and numbered.

[0086] 2. Method

[0087] Concrete steps refer to embodiment 3.

[0088] 3. Results

[0089] Real-time quantitative PCR result display (see figure 2 ): The results of qRT-PCR amplification were stable, and the expression levels of TMED2, CCDC72 and ERC1 in the peripheral blood of postmenopausal osteoporosis were significantly different from those of healthy people. The former of the three genes was about 1.22% of the latter. times, 0.83 times and 1.39 times, the above results verified the results of high-throughput sequencing.

[0090] The present invention uses high-throughput sequencing to screen out postmenopausal osteoporosis-related genes TMED2, CCDC72 and ERC1, and further analy...

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Abstract

The invention relates to gene detection compositions and application thereof, in particular to application of TMED2, CCDC72 and ERC1 genes in detection of postmenopausal osteoporosis and clinical medication of Liuweidihuang pills. The inventor conducts high-throughput sequencing on peripheral blood samples before and after postmenopausal osteoporosis patient take the Liuweidihuang pills and control peripheral blood samples of healthy persons by a high-throughput sequencing technology; the results shows that candidate genes TMED2, CCDC72 and ERC1 have obvious change after the postmenopausal osteoporosis patients take the Liuweidihuang pills, and further analysis results of high-throughput data are verified by a molecular biology experiment. The invention provides gene compositions for clinically and accurately diagnosing postmenopausal osteoporosis and accurate medication and has a good market application value.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a gene detection composition and its application, and more specifically to the application of TMED2, CCDC72 and ERC1 gene composition in detecting postmenopausal osteoporosis and Liuwei Dihuang Pills in clinical medicine. Background technique [0002] The current research on the function of TMED2 mostly focuses on its participation in the vesicle transport process between the endoplasmic reticulum and the Golgi apparatus. In the past two years, TMED2 has been proved to be a specific transport receptor of G protein-coupled receptors (GPCRs), and controls the transport process in the biosynthetic pathway of GPCRs, and regulates GPCR signaling pathways in astrocytes. In addition, it has been reported that TMED2 is expressed in human placenta and choriocarcinoma cell lines throughout pregnancy, and mutations in TMED2 in mice lead to developmental defects in embryos and placenta, suggesting...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/178
Inventor 常鹏孙耀兰
Owner GUAN BOJIAN BIOTECH CO LTD
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