CN107164507A - The diagnostic tool of postmenopausal women's primary osteoporosis - Google Patents

Abstract

The invention discloses a diagnostic tool for postmenopausal women's primary osteoporosis. The diagnostic tool realizes the diagnostic purpose by detecting the DHRS12 gene and its expression product in blood. The research of the present invention proves that compared with normal postmenopausal women, the mRNA expression level of DHRS12 gene in the blood of postmenopausal women with primary osteoporosis is significantly decreased. According to the correlation between the DHRS12 gene and primary osteoporosis in postmenopausal women, a kit for diagnosing primary osteoporosis in postmenopausal women can be prepared, and the kit can be widely used clinically.

Description

Diagnostic tool for primary osteoporosis in postmenopausal women

technical field

The invention belongs to the field of diagnosis, relates to a diagnostic tool for primary osteoporosis in postmenopausal women, and also relates to the application of DHRS12 gene in blood in preparing the diagnostic tool for primary osteoporosis in postmenopausal women.

Background technique

Osteoporosis (OP) is a systemic bone disease characterized by low bone mass, destruction of bone microarchitecture, increased bone fragility, and susceptibility to fracture (WHO). In 2001, the National Institutes of Health (NIH) of the United States proposed that osteoporosis is a disease of the skeletal system characterized by decreased bone strength and increased risk of fracture. Bone strength reflects two main aspects of bones, namely bone mineral density and bone quality. .

The disease can occur in different genders and at any age, but it is more common in postmenopausal women and older men. Osteoporosis is divided into primary and secondary two categories. Primary osteoporosis is further divided into three types: postmenopausal osteoporosis (type I), senile osteoporosis (type II) and idiopathic osteoporosis (including juvenile type). Postmenopausal osteoporosis generally occurs in women within 5-10 years after menopause. It is caused by the decrease of estrogen synthesized by postmenopausal ovary. systemic disease. Postmenopausal women lose bone mineral density (BMD) rapidly, with an annual loss rate of 1.5%-2.5%, resulting in a sharp decline in bone mineral density, making postmenopausal women a high-risk population for osteoporosis.

Individuals with the following high-risk factors are predisposed to developing postmenopausal osteoporosis:

Estrogen deficiency: Estrogen deficiency is considered to be the main factor causing postmenopausal osteoporosis. The postmenopausal ovarian function declines, the level of estrogen secretion decreases, and at the same time, the intake of vitamin D and calcium is insufficient, resulting in secondary osteoporosis. Bone loss.

Genetic factors: Osteoporosis is more common in whites, followed by yellows, and the least in blacks. Bone mineral density is the main index for diagnosing osteoporosis. The value of bone mineral density is mainly determined by genetic factors, followed by environmental factors.

Nutritional factors: Calcium deficiency, it has been found that calcium intake in adolescence is directly related to peak bone mass in adults. 3/4 of people with low calcium diet (<10mg·kg ^-1 ·d ^-1 ) suffer from osteoporosis, while 1/4 of those with high calcium diet (10mg·kg ^-1 ·d ^-1 ) suffer from osteoporosis Osteoporosis; long-term lack of protein nutrition, resulting in decreased plasma protein, insufficient bone matrix synthesis, and lagging new bone formation, as if accompanied by calcium deficiency, osteoporosis will accelerate; vitamin C deficiency is the inability of bone matrix hydroxyproline synthesis Lack of, such as lack of bone matrix synthesis can be reduced.

Disuse factors: Muscle is a kind of mechanical force on bone tissue. If muscle is developed, the bone will be strong and the bone density will be high. The elderly have reduced activity, weakened muscle strength, reduced mechanical stimulation, and reduced bone mass. At the same time, the weakening of muscle strength and coordination disorders make the elderly more likely to fall, and when accompanied by bone loss, fractures have already occurred.

Other factors: Alcoholism, smoking, excessive coffee and caffeine intake, and heparin use are all risk factors for this disease. Alcoholism can affect calcium absorption, and alcohol also acts directly on osteoblasts, inhibiting bone formation. Excessive caffeine intake is the loss of urinary calcium and endogenous fecal calcium: heparin can cause osteoporosis, general research shows that 60% of patients who receive more than 1500U of heparin per day develop into osteoporosis.

In addition, increasing age, use of corticosteroids and anticonvulsant drugs, family history of osteoporosis or specific genes affecting bone mass, early menopause or bilateral oophorectomy before menopause are also high risk factors for postmenopausal osteoporosis .

Postmenopausal osteoporosis may have no symptoms in the early stage. Bone loss occurs unconsciously, and it is not realized until a fracture occurs. It is also called the "silent killer". Because there are no typical clinical symptoms in the early stage, it is often It cannot attract enough attention from doctors and patients, so that many patients can only helplessly face the long road from illness to recovery.

To diagnose osteoporosis, bone density must first be measured. Bone density actually indicates the health of the bones, or conversely, the aging of the bones. Commonly used detection methods:

X-ray film method: the earliest use history, but due to radioactivity, its test results cannot be quantified, and it has been gradually replaced.

Single photon detector: low cost, convenient, small radiation and safe. However, it is limited due to the inability to measure the trunk bone and the inability to distinguish cortical bone, cancellous bone, and soft tissue, and has gradually been replaced.

Quantitative ultrasound: cheap, convenient, and no radiation damage. It is suitable for bone status census work of people of all ages. It can not only detect bone density, but also understand the quality of bone, which can reflect the microstructure and elasticity of bone. However, it is currently only used for the calcaneus, tibia and phalanges, and cannot be used for the detection of whole body bones.

Dual-energy X-ray absorptiometry: It is the most ideal measurement method at present, and it is the gold standard for diagnosing osteoporosis. Expensive and therefore economically burdensome for diagnosis in osteoporosis patients. Therefore, finding a method for diagnosing osteoporosis with high sensitivity, high accuracy and reasonable price is an urgent problem to be solved.

In recent years, genetic factors have become one of the most active research fields in bone biology, and many genes have been identified as candidate genes regulating ancient BMD. Most of these candidate genes are genes that affect bone metabolism. Such as vitamin D receptor (VDR), lipoprotein receptor-related gene 5 (LRP5), type I collagen (COLIA1), estrogen receptor alpha, transforming growth factor TGF beta 1, bone morphogenetic proteins (BMPs), Runx2, Osterix (Osx), Sclerostin, TCIRG1, IGF-1, IL-1 (osteoporosis, relaxation, Chinese Journal of Osteoporosis, October 2010, 802-813).此外，已经有相当数量的专利申请涉及骨质疏松症的基因诊断，如申请号为：201610272604.0、201610922798.4、201510628024.6、201510628081.4、201510725408.X、201510628042.4、201610530383.2、201510629348.1、201510627056.4、201510627060.0、201610555353.7的专利。 It can be seen that the use of genes to diagnose osteoporosis has become the development trend of early diagnosis of osteoporosis in the future.

Contents of the invention

The present invention finds for the first time that the content of DHRS12 (Dehydrogenase/reductase 12) gene in the blood of patients with primary osteoporosis in postmenopausal women is much lower than that of normal people. A method for osteoporosis, whereby a tool for diagnosing primary osteoporosis in postmenopausal women could be developed.

Specifically, the present invention provides the application of the product for detecting the expression of DHRS12 gene in the preparation of tools for diagnosing primary osteoporosis in postmenopausal women.

Further, the detection products mentioned above include: detection of the expression level of DHRS12 gene by RT-PCR, real-time quantitative PCR, immunoassay, in situ hybridization, chip or high-throughput sequencing platform to diagnose primary osteoarthritis in postmenopausal women Porosis products.

Further, the product for diagnosing primary osteoporosis in postmenopausal women by RT-PCR includes at least a pair of primers for specific amplification of the DHRS12 gene; the diagnosis of primary osteoporosis in postmenopausal women by real-time quantitative PCR The product includes at least one pair of primers for specific amplification of the DHRS12 gene; the product for diagnosing primary osteoporosis in postmenopausal women by immunoassay includes: an antibody specifically binding to the DHRS12 protein; the in situ hybridization diagnosis The products for primary osteoporosis in postmenopausal women include: probes hybridized with the nucleic acid sequence of the DHRS12 gene; the products for diagnosing primary osteoporosis in postmenopausal women with chips include: protein chips and gene chips; Wherein, the protein chip includes an antibody specifically combined with the DHRS12 protein, and the gene chip includes a probe hybridized with the nucleic acid sequence of the DHRS12 gene.

In a specific embodiment of the present invention, the product for diagnosing primary osteoporosis in postmenopausal women by real-time quantitative PCR includes at least a pair of sequences of primers for specific amplification of DHRS12 gene such as SEQ ID NO.3 and SEQ ID NO .4 shown.

Preferably, the diagnostic tools include chips, kits, test strips or high-throughput sequencing platforms. Among them, the high-throughput sequencing platform is a special diagnostic tool, and products for detecting DHRS12 gene expression can be applied to this platform to detect the expression of DHRS12 gene. With the development of high-throughput sequencing technology, the construction of a person's gene expression profile will become a very convenient task. By comparing the gene expression profiles of disease patients and normal people, it is easy to analyze which gene abnormality is related to the disease. Therefore, it is known in high-throughput sequencing that the abnormality of the DHRS12 gene is related to primary osteoporosis in postmenopausal women, which also belongs to the use of the DHRS12 gene, and is also within the protection scope of the present invention.

The invention also provides a tool for diagnosing primary osteoporosis in postmenopausal women, and the product includes a chip, a kit, a test strip, or a high-throughput sequencing platform.

Wherein, the chip includes a gene chip and a protein chip; the gene chip includes a solid phase carrier and an oligonucleotide probe fixed on the solid phase carrier, and the oligonucleotide probe includes The oligonucleotide probe for the DHRS12 gene; the protein chip includes a solid phase carrier and the specific antibody of the DHRS12 protein immobilized on the solid phase carrier; the gene chip can be used to detect multiple genes including the DHRS12 gene (eg, expression levels of multiple genes associated with primary osteoporosis in postmenopausal women). The protein chip can be used to detect the expression levels of multiple proteins including DHRS12 protein (for example, multiple proteins related to primary osteoporosis in postmenopausal women). By simultaneously detecting multiple markers of primary osteoporosis in postmenopausal women, the accuracy of diagnosis of primary osteoporosis in postmenopausal women can be greatly improved.

Wherein, the kit includes a gene detection kit and a protein immune detection kit; the gene detection kit includes reagents for detecting DHRS12 gene transcription level; the protein immune detection kit includes a specific antibody for DHRS12 protein. Further, the reagents include the reagents required in the process of detecting the expression level of DHRS12 gene by using RT-PCR, real-time quantitative PCR, immunoassay, in situ hybridization or chip method. Preferably, the reagents include primers and/or probes for the DHRS12 gene. Primers and probes that can be used to detect the expression level of the DHRS12 gene can be easily designed according to the nucleotide sequence information of the DHRS12 gene.

The high-throughput sequencing platform includes reagents for detecting the expression level of the DHRS12 gene.

The test paper includes a test paper carrier and an oligonucleotide fixed on the test paper carrier, and the oligonucleotide can detect the transcription level of the DHRS12 gene.

The probe hybridizing with the nucleic acid sequence of the DHRS12 gene can be DNA, RNA, DNA-RNA chimera, PNA or other derivatives. The length of the probe is not limited, as long as it completes specific hybridization and specifically binds to the target nucleotide sequence, any length is acceptable. The probes can be as short as 25, 20, 15, 13 or 10 bases in length. Likewise, the probes can be as long as 60, 80, 100, 150, 300 base pairs or longer, or even the entire gene. Since different probe lengths have different effects on hybridization efficiency and signal specificity, the length of the probe is usually at least 14 base pairs, and the longest is generally no more than 30 base pairs. The optimal length of complementarity is 15-25 base pairs. The self-complementary sequence of the probe is preferably less than 4 base pairs, so as not to affect the hybridization efficiency.

Further, the specific antibody of DHRS12 protein includes monoclonal antibody and polyclonal antibody. The specific antibody of the DHRS12 protein includes the complete antibody molecule, any fragment or modification of the antibody (for example, chimeric antibody, scFv, Fab, F(ab') 2, Fv, etc. As long as the fragment can be retained with DHRS12 Protein-binding ability is enough. Preparation of antibodies for protein level is well known to those skilled in the art, and any method can be used in the present invention to prepare the antibodies.

In a specific embodiment of the present invention, the primer sequence for the DHRS12 gene is as follows: the sequence of the forward primer is shown in SEQ ID NO.3, and the sequence of the reverse primer is shown in SEQ ID NO.4.

The sources of the DHRS12 gene and its expression products used for diagnosing primary osteoporosis in postmenopausal women include but are not limited to blood, interstitial fluid, urine, saliva, spinal fluid and other body fluids from which genomic DNA can be obtained. In a specific embodiment of the present invention, the source of the DHRS12 gene and its expression product for diagnosing primary osteoporosis in postmenopausal women is blood. In a specific embodiment of the invention, the blood is obtained from postmenopausal women with primary osteoporosis and from women with normal bone density.

The specific sequence of the DHRS12 gene of the present invention can be found in GeneBank, an international public nucleic acid sequence database.

The coding sequence of the DHRS12 gene of the present invention includes any one of the following DNA molecules:

(1) the DNA sequence shown in SEQ ID NO.1 in the sequence listing;

(2) A DNA sequence that hybridizes to the DNA sequence defined in 1) under stringent conditions and encodes the same functional protein;

(3) A DNA molecule having 70%, preferably more than 90%, homology with the DNA sequence defined in (1) or (2), and encoding the same functional protein.

In a specific embodiment of the present invention, the coding sequence of the DHRS12 gene is the DNA sequence shown in SEQ ID NO.1.

In the context of the present invention, DHRS12 gene expression products include DHRS12 protein and partial peptides of DHRS12 protein. The partial peptide of the DHRS12 protein contains a functional domain related to primary osteoporosis in postmenopausal women.

"DHRS12 protein" includes DHRS12 protein and any functional equivalents of DHRS12 protein. The functional equivalents include DHRS12 protein conservative variant proteins, or active fragments thereof, or active derivatives thereof, allelic variants, natural mutants, induced mutants, DNA capable of binding to DHRS12 under high or low stringent conditions The protein encoded by the hybridized DNA.

Preferably, the DHRS12 protein is a protein having the following amino acid sequence:

(1) A protein consisting of the amino acid sequence shown in SEQ ID NO.2 in the sequence listing;

(2) The amino acid sequence shown in SEQ ID NO.2 is subjected to substitution and/or deletion and/or addition of one or several amino acid residues and has the same function as the amino acid sequence shown in SEQ ID NO.2. A protein derived from the amino acid sequence shown in ID NO.2. The number of substituted, deleted or added amino acids is usually 1-50, preferably 1-30, more preferably 1-20, most preferably 1-10.

(3) having at least 80% homology (also known as sequence identity) with the amino acid sequence shown in SEQ ID NO.2, more preferably at least about 90% to the amino acid sequence shown in SEQ ID NO.2 95% homology, usually 96%, 97%, 98%, 99% homology of amino acid sequence polypeptide.

In a specific embodiment of the present invention, the DHRS12 protein is a protein having the amino acid sequence shown in SEQ ID NO.2.

In general, it is known that the modification of one or more amino acids in a protein does not affect the function of the protein. Those skilled in the art will recognize that changes to single amino acids or small percentages of amino acids or individual additions, deletions, insertions, substitutions to an amino acid sequence are conservative modifications where changes to a protein result in a protein with similar function. Conservative substitution tables providing functionally similar amino acids are well known in the art.

An example of a protein modified by adding an amino acid or amino acid residues is a fusion protein of the DHRS12 protein. There is no limitation on the peptide or protein fused with the DHRS12 protein, as long as the resulting fusion protein retains the biological activity of the DHRS12 protein.

The DHRS12 protein of the present invention also includes non-conservative modifications to the amino acid sequence shown in SEQ ID NO.2, as long as the modified protein can still retain the biological activity of the DHRS12 protein. The number of amino acids mutated in such modified proteins is usually 10 or less, such as 6 or less, such as 3 or less.

In the context of the present invention, "diagnosing primary osteoporosis in postmenopausal women" includes judging whether postmenopausal women already suffer from primary osteoporosis, and also includes judging whether postmenopausal women have primary osteoporosis It also includes predicting the prognosis of patients with primary osteoporosis in postmenopausal women, and also includes judging whether postmenopausal women with primary osteoporosis have relapsed after treatment.

Description of drawings

Figure 1 shows the difference in expression of DHRS12 gene detected by QPCR in postmenopausal women with primary osteoporosis and normal menopausal women;

Figure 2 shows the difference in expression of the DHRS12 gene detected by sequencing between postmenopausal women with primary osteoporosis and normal menopausal women.

specific implementation

The present invention will be described in further detail below in conjunction with the accompanying drawings and embodiments. The following examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental method that does not indicate specific condition in the embodiment, usually according to conventional conditions, such as Sambrook et al., molecular cloning: the conditions described in the laboratory handbook (New York: Cold Spring Harbor Laboratory Press, 1989), or according to the manufacturer's suggestion conditions of.

Example 1 Screening of genes differentially expressed in postmenopausal women with primary osteoporosis and normal menopausal women

1. Research object:

Three postmenopausal women with primary osteoporosis, aged 81, 68, and 79 years old, and two normal postmenopausal women, aged 67 and 68, were selected.

Inclusion criteria for postmenopausal women with primary osteoporosis: Han postmenopausal women who have menopause for more than 12 months, and all patients have informed consent;

Exclusion criteria for patients with primary osteoporosis in postmenopausal women include ① Previously received anti-resorptive drugs (including calcitonin, bisphosphonates, estrogen, etc.) or bone-promoting drugs (such as parathyroid hormone, etc.) Treatment (except for those who simply use calcium and/or vitamin D); ② long-term use of glucocorticoids; ③ combined with malignant tumors, bone metastases, and other endocrine and bone metabolism history (such as rheumatoid arthritis, hyperthyroidism, Adrenal gland disease, etc.); ④fresh fractures of the spine or hip or hip joint replacement; ⑤severe liver and kidney disease, long-term bed rest ≥ 3 months; ⑥mental disorders, cognitive impairments.

2. Total RNA extraction from blood

The blood total RNA was extracted using Bolinko TRIpure LS Reagent blood (blood sample) RNA extraction reagent.

The basic steps:

(1) Add TRIpure LS reagent and blood in a volume ratio of 3:1 and mix well;

(2) adding chloroform to help the organic phase and the aqueous phase delamination;

(3) isopropanol precipitation RNA;

(4) Rinse the RNA precipitation;

(5) RNase-free H ₂ O redissolves the RNA pellet.

3. Quality analysis of RNA samples

The concentration and purity of the extracted RNA were detected by Nanodrop2000, the RNA integrity was detected by agarose gel electrophoresis, and the RIN value was determined by Agilent2100. A single library construction requires a total amount of RNA of 5 μg, a concentration ≥ 200 ng/μL, and an OD260/280 between 1.8 and 2.2.

5. Fragmented RNA

The Illumina platform is for sequencing short sequence fragments, and the average length of mRNA may reach several kb, so it needs to be randomly interrupted. Using metal ions, RNA can be randomly broken into small fragments of about 200bp.

6. Inversion synthesis of cDNA

Under the action of reverse transcriptase, random primers are used to reversely synthesize one-strand cDNA using mRNA as a template. When performing second-strand synthesis, dUTP is used instead of dTTP in the dNTPs reagent, so that the bases in the second strand of cDNA contain A/U/ C/G.

7. Connect the adapter

The double-stranded cDNA structure is a cohesive end. Add End Repair Mix to make it a blunt end, and then add an A base at the 3' end to connect the Y-shaped adapter.

8. UNG enzyme digests the second strand of cDNA

Before PCR amplification, the second strand of cDNA was digested with UNG enzyme, so that only the first strand of cDNA was included in the library.

9. Illumina x-ten on-machine sequencing

Illumina x-ten sequencing platform for 2*150bp sequencing.

10. Bioinformatics analysis

The rawdata analysis process after the sequencing data is obtained is as follows:

(1) Use cutadapt to trim the 5' and 3' segments of the reads, trim the bases with a quality <20, and delete the reads with N greater than 10%;

(2) tophat is compared to the reference genome. The reference genome version used is GRCh38.p7, and the fasta and gff files are downloaded from NCBI;

(3) cuffquant quantifies the expression level of mRNA and normalizes the output;

(4) Use the DEGseq package in the R environment to compare the mRNA expression differences between the control group and the disease group. Significantly different mRNA screening conditions: p-value<0.05.

11. Results

The results of RNA-seq showed (as shown in Figure 2) that compared with normal postmenopausal women, the mRNA level of DHRS12 gene in the blood of postmenopausal women with primary osteoporosis decreased significantly, and the difference was statistically significant (P<0.05 ).

Example 2 QPCR experiment verification of genes differentially expressed in postmenopausal women with primary osteoporosis and normal menopausal women

1. Research object:

Select 30 postmenopausal women with primary osteoporosis and 30 normal postmenopausal women, aged 65-82 years.

Inclusion criteria for postmenopausal women with primary osteoporosis: Han postmenopausal women who have menopause for more than 12 months, and all patients have informed consent;

Exclusion criteria for patients with primary osteoporosis in postmenopausal women include ① Previously received anti-resorptive drugs (including calcitonin, bisphosphonates, estrogen, etc.) or bone-promoting drugs (such as parathyroid hormone, etc.) Treatment (except for those who simply use calcium and/or vitamin D); ② long-term use of glucocorticoids; ③ combined with malignant tumors, bone metastases, and other endocrine and bone metabolism history (such as rheumatoid arthritis, hyperthyroidism, Adrenal gland disease, etc.); ④fresh fractures of the spine or hip or hip joint replacement; ⑤severe liver and kidney disease, long-term bed rest ≥ 3 months; ⑥mental disorders, cognitive impairments.

2. Total RNA extraction from blood

The blood total RNA was extracted using Bolinko TRIpure LS Reagent blood (blood sample) RNA extraction reagent.

The basic steps:

(1) Add TRIpure LS reagent and blood in a volume ratio of 3:1 and mix well;

(2) adding chloroform to help the organic phase and the aqueous phase delamination;

(3) isopropanol precipitation RNA;

(4) Rinse the RNA precipitation;

(5) RNase-free H ₂ O redissolves the RNA pellet.

3. Reverse transcription

1 μg of total RNA was reverse-transcribed to synthesize cDNA using reverse transcription buffer. Use 25μl reaction system, take 1μg total RNA for each sample as template RNA, add the following components to the PCR tube respectively: DEPC water, 5× reverse transcription buffer, 10mmol/L dNTP, 0.1mmol/l DTT, 30μmmol/l Oligo dT, 200 U/μl M-MLV, template RNA. Incubate at 42°C for 1h, then centrifuge briefly at 72°C for 10min.

4. QPCR

A 25 μl reaction system was used, three parallel tubes were set up for each sample, and all amplification reactions were repeated more than three times to ensure the reliability of the results.

Prepare the following reaction system: SYBR Green polymerase chain reaction system 12.5 μl, forward primer (5 μM/μl) 1 μl, reverse primer (5 μM/μl) 1 μl, template cDNA 2.0 μl, enzyme-free water 8.5 μl.

The primer sequences are as follows:

The forward primer sequence for amplifying the DHRS12 gene is 5'-CAGTCCGAAAGAACACCAT-3' (SEQ ID NO.3), and the reverse primer sequence is 5'-CTCCGTCAGAACCACTTG-3' (SEQ ID NO.4);

The sequence of the forward primer for amplifying the GAPDH gene is 5'-TTTAACTCTGGTAAAGTGGATAT-3' (SEQ ID NO.5), and the sequence of the reverse primer is 5'-GGTGGAATCATATTGGAACA-3' (SEQ ID NO.6).

Amplification program: 95°C for 5min, (95°C for 10s, 60°C for 60s)*45 cycles. to SYBR

Green was used as a fluorescent marker, and the PCR reaction was carried out on a Light Cycler fluorescence real-time quantitative PCR instrument. The target band was determined by melting curve analysis and electrophoresis, and the relative quantification was carried out by the ΔΔCT method.

5. Results

The results are shown in Figure 1. Compared with normal postmenopausal women, the mRNA level of DHRS12 gene in the blood of postmenopausal women with primary osteoporosis decreased significantly, and the difference was statistically significant (P<0.05). The results were the same as those of RNA- seq experiment.

The description of the above embodiments is only for understanding the method and core idea of the present invention. It should be pointed out that for those skilled in the art, without departing from the principles of the present invention, some improvements and modifications can be made to the present invention, and these improvements and modifications will also fall within the protection scope of the claims of the present invention.

SEQUENCE LISTING

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Claims (10)

1. detect the product of DHRS12 gene expressions in the instrument for preparing diagnosis postmenopausal women's primary osteoporosis Using.

2. application according to claim 1, it is characterised in that the product includes：By RT-PCR, real-time quantitative PCR, Immune detection, in situ hybridization, the expression of chip or high-flux sequence detection of platform DHRS12 genes are to diagnose post menopausal woman The product of female's primary osteoporosis.

3. application according to claim 2, it is characterised in that described to diagnose postmenopausal women's primary sclerotin with RT-PCR The product of osteoporosis at least includes the primer of a pair of specific amplified DHRS12 genes；It is described to diagnose post menopausal with real-time quantitative PCR The product of women's primary osteoporosis at least includes the primer of a pair of specific amplified DHRS12 genes；It is described to use immune detection The product of diagnosis postmenopausal women's primary osteoporosis includes：The antibody combined with DHRS12 protein-specifics；It is described to use The product of in situ hybridization diagnosis postmenopausal women's primary osteoporosis includes：With the nucleic acid array hybridizing of DHRS12 genes Probe；The product for diagnosing postmenopausal women's primary osteoporosis with chip includes：Protein chip and genetic chip；Its In, protein chip includes the antibody combined with DHRS12 protein-specifics, and genetic chip includes the nucleic acid sequence with DHRS12 genes Arrange the probe of hybridization.

4. application according to claim 3, it is characterised in that described primary with real-time quantitative PCR diagnosis postmenopausal women Primer such as SEQ ID NO.3 and the SEQ ID for a pair of specific amplified DHRS12 genes that the product of property osteoporosis at least includes Shown in NO.4.

5. a kind of instrument for being used to diagnose postmenopausal women's primary osteoporosis, it is characterised in that the instrument can lead to The expression of DHRS12 genes in detection sample is crossed to diagnose postmenopausal women's primary osteoporosis.

6. instrument according to claim 5, it is characterised in that the instrument includes chip, kit, test paper or high flux Microarray dataset.

7. instrument according to claim 6, it is characterised in that the chip includes genetic chip, protein-chip；It is described Genetic chip includes solid phase carrier and is fixed on the oligonucleotide probe of solid phase carrier, and the oligonucleotide probe includes being used for Detect the oligonucleotide probe for DHRS12 genes of DHRS12 gene transcription levels；The protein-chip is carried including solid phase Body and be fixed on solid phase carrier DHRS12 albumen specific antibody；The kit includes gene detecting kit and egg White immunity detection reagent；The gene detecting kit includes the reagent for being used to detect DHRS12 gene transcription levels；It is described Protein immunization detection kit includes the specific antibody of DHRS12 albumen；The test paper includes being used to detect that DHRS12 genes turn The reagent of record level；The high-flux sequence platform includes the reagent for being used to detect DHRS12 gene transcription levels.

8. instrument according to claim 7, it is characterised in that the reagent of the detection DHRS12 gene transcription levels includes For the primer and/or probe of DHRS12 genes.

9. instrument according to claim 8, its spy is characterised by, the primer sequence for DHRS12 genes is as follows： Forward primer sequence is as shown in SEQ ID NO.3, and reverse primer is as shown in SEQ ID NO.4.

10. the instrument according to any one of claim 5-9, it is characterised in that the sample is blood.