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Gene detection compositions and application thereof

A composition and gene technology, applied in the field of biomedicine, can solve problems such as unproven correlation, and achieve the effect of simplifying the process of quantitative detection, strong selectivity and strong specificity

Active Publication Date: 2018-05-29
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)

Method used

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  • Gene detection compositions and application thereof
  • Gene detection compositions and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1 High-throughput sequencing and analysis

[0050] Collect the peripheral blood samples of 3 cases of postmenopausal osteoporosis before and after taking Liuwei Dihuang Pills and 3 cases of healthy control peripheral blood samples for RNA extraction, agarose gel electrophoresis after RNA extraction, and preliminary extraction from the electrophoresis results Whether the quality of the RNA sample is qualified or not, can it be used for further transcriptome analysis. Furthermore, the extraction of RNA samples was detected by NanoDrop1000 spectrophotometer. The sample requirements for RNA-seq sequencing: OD260 / OD280 is 1.8-2.2.

[0051] The sequencing platform is Illumina's HiSeq 2500 high-throughput sequencing platform, 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 base quality value distribution and quality value Location distribution, G...

Embodiment 2

[0052] Example 2 The expression of TMED2, CCDC72 and ERC1 genes in peripheral blood of postmenopausal patients with osteoporosis before and after taking Liuwei Dihuang Wan

[0053] 1. Materials and methods

[0054] 1. Material

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

[0056] 2. Method

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

[0058] use Reagent conducts sample RNA extraction, and the experimental operation is carried out according to the product manual.

[0059] RNA quality criterion: the OD260 / OD280 value of the RNA sample is between 1.8-2.2; the total RNA electrophoresis pattern has clear 28S, 18S bands; the electrophoresis pattern after 1 hour of incubation a...

Embodiment 3

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

[0084] 1. Material

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

[0086] 2. Method

[0087] Refer to Example 3 for specific steps.

[0088] 3. Results

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

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

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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 the TMED2, CCDC72 and ERC1 gene composition in detecting postmenopausal osteoporosis and Liuwei Dihuang Pills for clinical medication. Background technique [0002] The current research on the function of TMED2 mostly focuses on its involvement in the vesicle transport process between the endoplasmic reticulum and the Golgi apparatus. In the past two years, TMED2 has been proven to be a specific transport receptor for G protein-coupled receptors (GPCRs), and controls the transport process in the GPCR biosynthetic pathway, and regulates the GPCR signaling pathway in astrocytes. In addition, there are reports in the literature that TMED2 is expressed in human placenta and choriocarcinoma cell lines throughout pregnancy. Mutation of TMED2 in mice leads to embryonic and placental development defects, s...

Claims

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Application Information

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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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