Serum microRNA diagnostic marker for osteoportic fractures in old people and diagnostic kit thereof

A diagnostic kit and diagnostic marker technology, applied in the field of serum microRNA diagnostic markers, can solve problems such as poor sensitivity, and achieve the effects of good stability, simple diagnostic steps, and improved accuracy and sensitivity

Inactive Publication Date: 2017-04-26
NORTHWESTERN POLYTECHNICAL UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In order to overcome the insufficient sensitivity of the existing serum microRNA diagnostic kits for senile osteoporotic fractures, the present invention provides a serum microRNA diagnostic marker for senile osteoporotic fractures and its diagnostic kit

Method used

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  • Serum microRNA diagnostic marker for osteoportic fractures in old people and diagnostic kit thereof
  • Serum microRNA diagnostic marker for osteoportic fractures in old people and diagnostic kit thereof
  • Serum microRNA diagnostic marker for osteoportic fractures in old people and diagnostic kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1: Screening microRNAs related to osteoporotic fractures in bone tissue.

[0028] 1. Collection of bone tissue samples from elderly patients with osteoporotic fractures.

[0029] From July 2014 to the present, the project team has collected primary knee joint disease, 111 cases (including male and female) of bone tissue were removed during surgery in patients with lumbar fractures, lumbar disc herniation, and hip / femur fractures, and detailed pathological information was obtained. Some patients were interviewed to obtain past medical history and treatment history and other information. After the samples were properly stored at -80°C, the samples were screened according to the inclusion criteria against the patient's medical records to obtain samples that could be used for bone tissue RNA extraction experiments.

[0030] 2. Chip experiment of microRNA expression profiling in bone tissue of patients with osteoporotic fractures.

[0031] Based on the bone tissue...

Embodiment 2

[0039] Example 2: Collection and preparation of serum clinical samples.

[0040] 1. Objects and methods.

[0041] 1.1 Samples and sources.

[0042] After obtaining the informed consent of the patients, blood was collected from 30 healthy people with BMD>-2.5 without osteoporosis and 30 patients with osteoporosis with BMD<-2.5 in Xi'an Honghui Hospital. After blood collection, record the detailed information of the patient's medical records, including past disease history, treatment history, and the latest diagnostic information such as T Score. And record the patient's age, ethnicity, education, work and other personal information, as well as drinking, smoking, diet and other necessary living habits information.

[0043] Exclusion criteria: under the age of 50, suffering from cardiovascular disease, cancer, rheumatoid arthritis and other hereditary and nutritional diseases that may cause skeletal system lesions; lung disease caused by long-term smoking; liver disease caused ...

Embodiment 3

[0050] Embodiment 3: Quantitative detection of microRNA in normal serum samples and osteoporosis serum samples.

[0051] 1. Extraction of microRNA in serum.

[0052] The microRNA extraction kit in serum was purchased from Tiangen Biochemical Technology Co., Ltd. The concentration of the extracted RNA was measured by using a micro-ultraviolet spectrophotometer, and the OD value of 260 / 280nm was obtained.

[0053] 2. Quantitative detection of microRNA in serum samples.

[0054] 2.1 Reverse transcription of RNA to cDNA.

[0055] Prepare the reaction solution according to the following system (the reaction solution is prepared on ice) as follows:

[0056] Table 1: microRNA reverse transcription system

[0057] 5×PrimeScript Buffer(for Real time) 2μl PrimeScript RT Enzyme Mix I 0.5μl Specific microRNA primer (50μM) 0.5μl Random 6mers (100μM) 0.5μl Total RNA 0.5μg RNase Free dH2O up to 10μl

[0058] The reverse transcription rea...

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Abstract

The invention discloses a serum microRNA diagnostic marker for osteoportic fractures in old people and a diagnostic kit thereof to overcome the technical problem of poor sensitivity of conventional serum microRNA diagnostic kits for osteoportic fractures in old people. According to a technical scheme in the invention, the microRNA biomarker is one selected from a group consisting of hsa-miR-497-5p, hsa-miR-1290, hsa-miR-4299, hsa-miR-181c-5p, hsa-miR-320a and hsa-miR-204-3p, or a combination of hsa-miR-181c-5p and hsa-miR-4299, or a combination of hsa-miR-497-5p and hsa-miR-1290. The marker and primers thereof can be applied to the diagnostic kit; and the diagnostic kit has high sensitivity when applied to diagnosis of osteoportic fractures in old people.

Description

technical field [0001] The invention relates to a serum microRNA diagnostic marker for senile osteoporotic fractures, and also relates to a serum microRNA diagnostic kit for senile osteoporotic fractures. Background technique [0002] Senile bone degenerative changes often cause osteoporosis, and severe osteoporosis can often lead to fractures, which not only increases the economic burden of patients, but also reduces their quality of life and even shortens their life span. The methods currently used to assess the risk of osteoporotic fractures include bone density testing, FRAX, and OSTA. Although methods such as bone density and FRAX are currently the main methods for predicting the risk of osteoporotic fractures, their limitations as clinical detection are constantly emerging. Bone density testing relies on various X-ray equipment, which not only requires a certain amount of radiation, but also is difficult to effectively reduce the cost of testing for patients; although...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/113
CPCC12Q1/6883C12Q2600/158C12Q2600/166C12Q2600/178
Inventor 骞爱荣胡丽芳马剑华李思宇赵帆陈志浩李迪杰陈楚杨团民印崇苏佩红马小莉张琰仇伍霞张茹王牌杨超飞
Owner NORTHWESTERN POLYTECHNICAL UNIV
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