CN113817818A - Tool for diagnosing allergic airway inflammation - Google Patents
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Abstract
The invention belongs to the field of biomedicine, and particularly relates to a tool for diagnosing allergic airway inflammation. The invention studies the diagnostic value of LINC02579 as a marker in allergic airway inflammation. Provides a new method for diagnosing allergic airway inflammation and has important clinical significance and application prospect.
Description
Technical Field
The invention belongs to the field of biomedicine, and particularly relates to a tool for diagnosing allergic airway inflammation.
Background
Allergic Rhinitis-Asthma Syndrome (Combined Allergic Rhinitis and arthritis Syndrome, CARAS) is a newly defined disease, which is a new disease name proposed by the World Allergy Organization (WAO) in 2004, and is defined as the simultaneous clinical or subclinical upper airway allergy (Allergic Rhinitis) and Allergic symptoms of the lower airway (bronchial Asthma), and the upper airway and lower airway Allergic symptoms often exist simultaneously. Recent studies indicate that Allergic Rhinitis (AR) and Bronchial Asthma (BA) are very similar in etiology, pathology, pathogenesis and the like, a plurality of patients suffer from AR and BA sequentially or simultaneously, the correlation of the two diseases has attracted wide attention of scholars at home and abroad, and in 2001, the World Health Organization (WHO) particularly issues Allergic rhinitis and the influence thereof on asthma for the two diseases (Allergic rhinitis and its impact on asthma), and proposes the concept of 'one respiratory tract and one disease'. Due to the close relationship between allergic rhinitis and asthma, the World Allergy Organization (WAO) and the following journal of Allergy & Clinical Immunology International and International Archives of Allergy and Immunology propose the use of the term allergic rhinitis-asthma syndrome according to the reader's feedback. At present, CARAS diagnosis is combined diagnosis of AR and BA, the diagnosis of allergic rhinitis in China at 2010 adopts the guidance for diagnosis and treatment of allergic rhinitis published by the editorial committee of journal of otolaryngology and neck surgery of China, and the diagnosis of bronchial asthma adopts the guidance for prevention and treatment of bronchial asthma (the definition, diagnosis, treatment and management scheme of bronchial asthma) published by the asthma institute of respiratory disease of Chinese medical society in 2008.
With the development of high throughput transcriptome sequencing technology, researchers have found that more than 90% of transcriptomes in the human genome are non-coding RNAs, of which micrornas (mirnas) have been thought to be involved in a variety of biological processes, including the development and progression of cancer. With the continued discovery of mirnas associated with cervical cancer, researchers have focused on another non-coding RNA, Long non-coding RNA (LncRNAs). LncRNAs are more than 200 nucleotide units in length and have no or only limited protein coding capacity. LncRNAs have the functions of regulating chromatin and regulating genes according to the positions in cells. LncRNAs can be stably expressed in cell lines and tissues, and have relatively stable properties in various body fluids, particularly LncRNAs in circulation can resist factors such as freezing, thawing or enzyme degradation, so that the research on the functions and the generation and development mechanisms of novel LncRNAs related to CARAS can provide scientific basis for searching high-specificity and sensitive biomarkers of CARAS and provide possibility for developing new more effective treatment.
Disclosure of Invention
The present invention relates to a tool for diagnosing allergic airway inflammation.
The present invention is based on the following findings of the inventors:
the invention adopts a SYBR Green I Real Time PCR method to detect the change condition of the transcription level of a target gene lncRNA in blood samples of a healthy control group and an allergic rhinitis-asthma syndrome patient group. The invention discovers for the first time that the expression of LINC02579 is reduced in a blood sample of a patient group with allergic rhinitis-asthma syndrome compared with a healthy control group, and suggests that LINC02579 can be used for diagnosing allergic airway inflammation.
Accordingly, in a first aspect, the present invention provides a reagent for determining the level or amount of a biomarker comprising LINC 02579.
In the present invention, biomarkers such as LINC02579(gene ID: 339807) include gene and mutations thereof. The term encompasses naturally occurring variants (e.g., splice variants) of the biomarker.
Further, the reagent includes a reagent for measuring the level of the biomarker or the amount thereof by microarray analysis, Polymerase Chain Reaction (PCR), reverse transcriptase polymerase chain reaction (RT-PCR), Northern blot, Southern blot, or Serial Analysis of Gene Expression (SAGE).
Further, the reagent comprises:
(1) a primer capable of specifically amplifying LINC 02579; or
(2) A probe capable of specifically hybridizing to LINC 02579.
Furthermore, the primer sequence is shown in SEQ ID NO. 5-6.
In a second aspect, the present invention provides a kit comprising the reagents of the first aspect of the invention.
In a third aspect, the present invention provides a chip comprising a reagent according to the first aspect of the invention.
In a fourth aspect, the invention provides a use as claimed in any one of:
(1) use of an agent according to the invention in a first aspect in the manufacture of a means for diagnosing allergic airway inflammation;
(2) use of a kit according to the invention in a second aspect for the manufacture of a means for diagnosing allergic airway inflammation;
(3) the use of a chip according to the third aspect of the invention for the manufacture of a tool for diagnosing allergic airway inflammation.
Further, the allergic airway inflammation includes allergic rhinitis, asthma, allergic rhinitis-asthma syndrome, preferably allergic rhinitis-asthma syndrome.
In a fifth aspect, the present invention provides a method of screening a candidate drug for treating allergic airway inflammation, comprising the steps of:
1) administering a drug to be tested to a subject in a test group, and detecting the expression level V1 of LINC02579 in a sample derived from said subject in the test group; in a control group, administering a blank control to the subject to be tested, and detecting the expression level V2 of LINC02579 in a sample derived from said subject in the control group;
2) comparing the level V1 and the level V2 detected in the previous step to determine whether the test compound is a candidate for treating allergic airway inflammation.
In a sixth aspect, the present invention provides a diagnostic system for performing a computer-implemented method for diagnosing a patient suspected of having an allergic airway inflammation, the computer performing steps comprising:
1) accepting input patient data comprising an expression level value for LINC02579 in a biological sample from the patient;
2) analyzing the level of the biomarker and comparing to a corresponding reference value range for the biomarker, if the patient's level of LINC02579 is lower than the reference value range for a non-diseased control subject, indicating that the patient has allergic airway inflammation;
3) displaying diagnostic information about the patient, wherein the diagnostic system comprises:
1) a storage component for storing data, wherein the storage component has instructions for determining a diagnosis of a subject stored therein;
2) a computer processor for processing data, wherein the computer processor is coupled to the storage component and configured to execute instructions stored in the storage component in order to receive patient data and analyze the patient data according to one or more algorithms; and
3) a display assembly for displaying diagnostic information about the patient;
further, the allergic airway inflammation includes allergic rhinitis, asthma, allergic rhinitis-asthma syndrome, preferably allergic rhinitis-asthma syndrome.
Drawings
FIG. 1 is an electrophoretogram of RNA;
FIG. 2 is a graph of internal reference GAPDH gene real-time amplification and product dissolution, wherein, a is a graph of GAPDH gene real-time amplification and B is a graph of product dissolution;
FIG. 3 is a LINC02579 gene real-time amplification graph and a product dissolution graph, wherein, a is a LINC02579 gene real-time amplification graph, and B is a product dissolution graph;
FIG. 4 is a bar graph of LINC02579 differential expression;
FIG. 5 is a ROC plot for LINC 02579.
Detailed Description
The terms used herein have the meanings commonly understood by those of ordinary skill in the art to which the present invention pertains. However, for a better understanding of the present invention, the definition and explanation of the related terms are as follows.
Terms such as "a," "an," and "the" are not intended to refer to only a single entity, but also include a class that can be described using specific examples.
In the present invention, the term "biomarker" (also referred to as "biological marker") refers to a measurable indicator of a biological state or condition of a subject. Such a biomarker may be any substance in a biological sample of the subject, such as a nucleic acid marker (e.g., LncRNA), so long as it is associated with a particular biological state or condition (e.g., disease) of the subject. Biomarkers are often measured and evaluated to detect normal biological processes, pathological processes, or pharmacological responses to therapeutic interventions, and are useful in many scientific fields.
In the present invention, biomarkers such as LINC02579(gene ID: 339807) include gene and mutations thereof. The term encompasses naturally occurring variants (e.g., splice variants) of the biomarker.
In the present invention, biomarkers can be detected and quantified by a variety of methods including, but not limited to, microarray analysis, Polymerase Chain Reaction (PCR), reverse transcriptase polymerase chain reaction (RT-PCR), Northern blot, Southern blot, and Serial Analysis of Gene Expression (SAGE). See, e.g., Draghicii "tools for Data analysis of DNA Microarrays (Data analysis tools for DNA microarray"), Chapman and Hall/CRC, 2003; simon et al Design and Analysis of DNA Microarray Studies (Design and Analysis of DNA Microarray investments), Springer, 2004; real-time PCR: current Technology and applications (Real-Time PCR: Current technologies and applications), Login, Edwards and Saunders, catalog Academic Press, 2009; burtin "A-Z of Quantitative PCR" (IUL Biotechnology, stage 5), International university Line, 2004; velculescu et al (1995) science 270: 484-; matsumura et al (2005) cytomicrobiology (cell. Microbiol.) 7: 11-18; serial Analysis of Gene Expression (SAGE): methods and protocols (Methods in Molecular Biology) (Serial Analysis of Gene Expression (SAGE): Methods and protocols (Methods in Molecular Biology)), (Humana Press, 2008); which is incorporated herein by reference in its entirety.
In the present invention, the term "biological sample" refers to a sample of tissue, cells or fluid isolated from a subject, including, but not limited to, samples such as blood, buffy coat, plasma, serum, blood cells (e.g., Peripheral Blood Mononuclear Cells (PBMCs), rod-shaped nuclear cells, neutrophils, monocytes or T cells), fecal matter, urine, bone marrow, bile, spinal fluid, lymph fluid, skin samples, external secretions of the skin, respiratory, intestinal, and genitourinary tracts, tears, saliva, milk, organs, biopsies, and in vitro cell culture components, including, but not limited to, conditioned media resulting from the growth of cells and tissue in culture, such as recombinant cells and cell components. In a specific embodiment of the present invention, the biological sample is blood.
In the present invention, the expression "a reagent for determining the level of a biomarker or the amount thereof" refers to a reagent that can be used to quantify or measure the level of a biomarker or the amount thereof in a sample. Such agents can be readily designed or obtained by conventional methods well known in the art based on the sequences of the biomarkers provided by the present invention. For example, such reagents include, but are not limited to, PCR primers that can be used to quantify or measure the level of a biomarker, or the amount thereof, by, for example, real-time PCR; probes useful for quantifying or measuring the level of a biomarker, or amount thereof, by, for example, quantitative Southern blotting; microarrays (e.g., gene chips) and the like that can be used to quantify or measure the levels of biomarkers or amounts thereof. In addition, second generation sequencing methods or third generation sequencing methods may also be used to quantify or measure the levels of biomarkers or their amounts, as is known in the art. Thus, such reagents may also be commercially available reagents for performing second generation sequencing methods or third generation sequencing methods.
According to the present invention, the term "second generation sequencing method" refers to a new generation of DNA sequencing methods developed in recent years, including, for example, Illumina GA, Roche 454, ABI Solid; and is different from conventional sequencing methods, such as Sanger sequencing methods. Second generation sequencing methods differ from traditional sequencing methods (such as Sanger sequencing methods) in that second generation sequencing methods analyze DNA sequences by sequencing-by-synthesis. The second generation sequencing method has the following advantageous aspects: 1) the cost is low and is 1 percent of that of the traditional sequencing method; 2) high throughput, simultaneous sequencing of multiple samples and one Solexa sequencing yields data of about 500 hundred million (50G) bases; 3) high precision (more than 98.4%) and effectively solves the problem of reading multiple repeated sequences. On the other hand, when the number of sequences to be sequenced has been predetermined, the high sequencing throughput in turn increases the sequencing depth of the sequences (e.g., each sequence may be sequenced multiple times), thereby ensuring the trustworthiness of the sequencing results.
According to the present invention, the term "third generation sequencing method" refers to a new generation of single molecule sequencing technology that has been recently developed. Third generation sequencing technologies offer advantageous aspects over current sequencing technologies, including (i) higher throughput; (ii) shorter turnaround times (e.g., the genomes of the animals after sequencing with high-fold coverage in minutes); (iii) longer sequencing length to enhance de novo assembly (de novo assembly) and enable direct detection of haplotypes (haplotypes) and even whole chromosome phasing (whole chromosome phasing); (iv) higher consistent accuracy to enable rare variation detection; (v) a small amount of starting material (theoretically only a single molecule is needed for sequencing); and (vi) low cost, where achieving high coverage sequencing of the human genome at prices below $ 100 has become a logical goal of society. For more details on third generation sequencing methods, see, e.g., Eric e.schadt et al, a window in third-generation sequencing, Human Molecular Genetics,2010, volume 19, Review Issue 2, R227-R240, incorporated herein by reference.
In the present invention, the expression "a primer capable of specifically amplifying" a specific nucleic acid or a specific sequence means that when used for amplification (e.g., PCR amplification), the primer specifically anneals to the specific nucleic acid or sequence and produces a unique amplification product (i.e., does not anneal to other nucleic acids or sequences or produce other byproducts).
In the present invention, the expression "a probe capable of specifically hybridizing to a specific nucleic acid or a specific sequence" means that the probe specifically anneals to and hybridizes to the specific nucleic acid hybridizing acid or sequence, but does not anneal to or hybridize to other nucleic acids or sequences, when used for hybridization or detection under stringent conditions.
It is common knowledge of the skilled person to design such primers or probes based on a specific sequence, such as LINC 02579. For example, such common general knowledge can be found in various textbooks (see, e.g., J.Sambrook et al, Molecular Cloning: Laboratory Manual, second edition, Cold Spring Harbor Laboratory Press, 1989; F.M. Ausubel et al, Short Protocols in Molecular Biology, third edition, John Wiley & Sons, Inc.; and many papers, such as Buck et al (1999), Lowe et al (1990), etc.
The phrase "differential expression" refers to a difference in the number and/or frequency of biomarkers present in a sample taken from a patient having, for example, allergic airway inflammation, as compared to a control subject or a subject not having the disease. For example, the biomarker may be a polynucleotide present at an elevated level or a reduced level in a sample from a patient with allergic airway inflammation as compared to a sample from a control subject. Alternatively, the biomarker may be a polynucleotide detected at a higher or lower frequency in a sample from a patient with allergic airway inflammation as compared to a sample from a control subject. Biomarkers can differ in number, frequency, or both.
A polynucleotide is differentially expressed between two samples if the amount of the polynucleotide in one sample is statistically significantly different from the amount of the polynucleotide in the other sample. For example, a polynucleotide is differentially expressed in two samples if it is present in an amount of at least about 120%, at least about 130%, at least about 150%, at least about 180%, at least about 200%, at least about 300%, at least about 500%, at least about 700%, at least about 900%, or at least about 1000% of the amount it is present in the other sample, or if it is detectable in one sample and not detectable in the other sample.
Alternatively or additionally, the polynucleotides are differentially expressed in the two groups of samples if the frequency of detection of the polynucleotides in the sample of a patient suffering from allergic airway inflammation is statistically significantly higher or lower than in the control sample. For example, a polynucleotide is differentially expressed in two sets of samples if it is detected that the polynucleotide is observed more frequently or less frequently in one set of samples than in another set of samples by at least about 120%, at least about 130%, at least about 150%, at least about 180%, at least about 200%, at least about 300%, at least about 500%, at least about 700%, at least about 900%, or at least about 1000%.
The terms "subject", "individual" and "patient" are used interchangeably herein and refer to any mammalian subject, particularly a human, in need of diagnosis, prognosis, treatment or therapy. Other subjects may include cows, dogs, cats, guinea pigs, rabbits, rats, mice, horses, and so forth. In some cases, the methods of the invention find application in the development of laboratory animals, veterinary applications, and animal disease models, including (but not limited to) rodents, including mice, rats, and hamsters, and primates.
The present invention will be described in further detail with reference to the accompanying drawings and examples. The following examples are intended to illustrate the invention only and are not intended to limit the scope of the invention. The experimental procedures, in which specific conditions are not specified in the examples, are generally carried out under conventional conditions or conditions recommended by the manufacturers.
Example 1Real Time PCR detection of changes in expression levels of target genes in blood samples from patients with allergic rhinitis-asthma syndrome
First, experiment purpose
Detecting the change condition of the transcription level of the target gene lncRNA in the blood sample of the patient with the allergic rhinitis-asthma syndrome by using a SYBR Green I Real Time PCR method.
Second, Experimental materials
1. Sample List
20 patients with allergic rhinitis-asthma syndrome and 20 normal persons were recruited in the study, clinical information is shown in Table 1, and blood samples were collected for analytical study.
TABLE 1 basic clinical information of the persons participating in the study
2. Experiment main reagent
TABLE 2 list of reagents used
3. Experiment main instrument
TABLE 3 List of instruments used
| Name of instrument | Instrument type | Manufacturer of the product |
| NanoVue Plus | 28956057 | BIOCHROM LTD |
| Fluorescent quantitative PCR instrument | ABI7300 | Applied Biosystems |
Third, Experimental methods
1. Primer design
1.1Real Time PCR detection of the target gene primer. The following primers were synthesized by Bomaide.
TABLE 4 primer sequences
2. Procedure of experiment
2.1 extraction of Total RNA from samples
Adding 0.75mL of lysis solution RLS to every 0.25mL of liquid sample (serum, plasma, cerebrospinal fluid, etc.), and blowing the liquid sample several times with a sample-adding gun to help lyse cells in the sample. Every 5 to 10 × 106At least 0.75ml of lysis solution was added to each cellAnd RLS. The final volume ratio of lysate RLS and liquid sample was always 3: 1.
② adding 0.75mL of lysis buffer RLS into an EP tube, then adding 0.25mL of blood sample, continuously shaking for 30s, mixing uniformly, and incubating for 10min at 15-30 ℃ to completely decompose the nucleoprotein body.
③ Per 0.75mL of lysis solution RLS, 0.2mL of chloroform was added thereto, and the mixture was shaken vigorously for 15 seconds and allowed to stand at room temperature for 5 min.
Fourthly, centrifuging for 10min at 12000rpm at 4 ℃, and dividing the sample into three layers: the lower organic phase, the middle layer and the upper colorless aqueous phase, and RNA is present in the upper aqueous phase. The volume of the aqueous layer was about 70% of the volume of RLS added, and the aqueous layer was transferred to a new tube for further processing.
Fifthly, adding 70% ethanol with 1 time volume (please check whether absolute ethanol is added or not), and reversing and mixing (at this time, precipitation may occur). The resulting solution, together with possible precipitates, is transferred to an adsorption column RA (which is fitted in a collection tube).
Sixthly, centrifuging at 12000rpm for 45s, discarding the waste liquid, and sleeving the adsorption column back to the collecting pipe again.
Seventhly, adding 0.5mL of deproteinized liquid RE, centrifuging at 12000rpm for 45s, and discarding waste liquid.
Add 0.5mL of rinsing solution RW (please check whether absolute ethanol is added first!), centrifuge at 12000rpm for 45s, and discard the waste liquid.
Ninthly, adding 0.5mL of rinsing liquid RW, centrifuging at 12000rpm for 45s, and discarding the waste liquid.
And (3) putting the adsorption column RA back into the pore collection tube, centrifuging at 13000rpm for 2min, and removing the rinsing liquid as much as possible so as to prevent residual ethanol in the rinsing liquid from inhibiting downstream reaction.
2.2 Synthesis of IncRNA cDNA by reverse transcription
Using FastQuant cDNA first strand synthesis kit (cat # KR106) to carry out lncRNA reverse transcription, firstly removing genomic DNA reaction, adding 5 XgDNA Buffer 2.0ul and TotalRNA 1ug into a test tube, adding RNase Free ddH2O to make the total volume to 10uL, heating in a water bath at 42 ℃ for 3min, and adding 10 Xfast RT Buffer 2.0uL, RT Enzyme Mix 1.0uL, FQ-RT Primer Mix 2.0uL, RNase Free ddH2O5.0 uL, mixing, adding into the test tube, mixing to give 20uL, heating in water bath at 42 deg.C for 15min and 95 deg.C for 3min, and storing at-20 deg.C or lower when the synthesized cDNA is required to be stored for a long time.
2.3 fluorescent quantitation of mRNA
2.3.1 Instrument and analytical method
Using ABI 7300 type fluorescent quantitative PCR instrument, adopting
The method performs a relatively quantitative analysis of the data. 2.3.2 the procedure is as follows:
(one) reaction system: amplification was carried out using SuperReal PreMix Plus (SYBR Green) (cat # FP205) and the experimental procedures were performed according to the product instructions. The RealTime reaction system is:
TABLE 5 RealTime reaction System
| Reagent | Amount of the composition used |
| 2×SuperReal PreMix Plus | 10μl |
| Upstream primer (10uM) | 0.6μl |
| Downstream primer (10uM) | 0.6μl |
| 50×ROX Reference Dye△ | 2μl |
| DNA template | 2ul |
| Sterilized distilled water | 4.8ul |
(II) the amplification procedure is as follows: 95 degrees 15min, (95 degrees 10sec, 55 degrees 30sec, 72 degrees 32sec) × 40 cycles, 95 degrees 15sec, 60 degrees 60sec, 95 degrees 15 sec).
(III) primer screening
Mixing cDNA of each sample, performing 10-fold gradient dilution by taking the cDNA as a template, taking 2 mu l of each diluted sample as the template, respectively amplifying by using a target gene primer and an internal reference gene primer, simultaneously performing melting curve analysis at 60-95 ℃, and performing primer screening according to the principle of high amplification efficiency and single peak of the melting curve.
(IV) sample RealTimePCR detection
After 10-fold dilution of each sample cDNA, 2. mu.l of each sample cDNA was used as a template, and the target gene primer and the reference gene primer were used for amplification, respectively (see Table five). At the same time, the dissolution curve analysis is carried out at 60-95 ℃.
TABLE 6 sample RealTimePCR detection design
| Form panel | Sample cDNA | Sample cDNA |
| Repeatedly detecting the number of |
3 | 3 |
| Primer and method for producing the same | Target gene primer | Internal reference gene primer |
(V) data statistics
Sorting original result ct values derived by running a program off-machine according to a sampling sequence to obtain three multi-hole original ct values of each gene of each sample, respectively calculating the average value of the three multi-hole ct values of a target gene and an internal reference gene in excel, respectively calculating the expression of the target gene relative to the internal reference gene in a control group (paracarcinoma tissue) and a test group (gastric cancer tissue), performing statistical analysis by adopting GraphPad software, and performing t test on the difference between the two.
Fourth, experimental results
RNA concentration detection results and 1.5% agarose RNA electrophoresis detection results
TABLE 7 RNA concentration and purity results
Note:
the RNA dissolved in water results in a low A260/280 ratio
A, the concentration does not reach the standard; b, unqualified A260/A280; c, unqualified electrophoretogram; h, qualified
Sample evaluation standard:
(1) the concentration is more than 30ng/ul
(2)1.8<A260/A280<2.0
(3) The electrophoretic pattern shows three distinct bands (the third band may not be visible)
TABLE 8 electrophoretic Loading
The RNA electrophoresis pattern is shown in FIG. 1, in which M represents DNA Marker: DM2000, from bottom to top in turn 100,250,500,750,1000 and 2000bp, wherein 750bp is a bright band.
2. Results and analysis of RealTimePCR detection for each sample
The real-time amplification curve and the dissolution curve of the sample amplification product of each sample are shown in FIGS. 2 and 3.
The statistical results are shown in fig. 4, and LINC02579 expression is down-regulated in the sample of the allergic rhinitis-asthma syndrome group compared to the healthy control group.
Example 2 diagnostic efficacy validation of LINC02579
SPSS software is used to draw a Receiver Operating Curve (ROC), AUC values, sensitivity and specificity are analyzed, and the individual diagnostic efficacy of the index is judged.
As shown in table 9 and fig. 5, LINC02579 has high diagnostic efficacy (AUC value of 0.898, sensitivity value of 0.850, specificity value of 0.800), suggesting that LINC02579 can be used for diagnosing allergic rhinitis-asthma syndrome.
TABLE 9 region under ROC Curve
The preferred embodiments of the present application have been described in detail with reference to the accompanying drawings, however, the present application is not limited to the details of the above embodiments, and various simple modifications can be made to the technical solution of the present application within the technical idea of the present application, and these simple modifications are all within the protection scope of the present application.
It should be noted that, in the foregoing embodiments, various features described in the above embodiments may be combined in any suitable manner, and in order to avoid unnecessary repetition, various possible combinations are not described in the present application.
In addition, any combination of the various embodiments of the present application is also possible, and the same should be considered as disclosed in the present application as long as it does not depart from the idea of the present application.
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Claims (10)
1. An agent for determining the level or amount of a biomarker, wherein the biomarker comprises LINC 02579.
2. The reagent of claim 1, wherein the reagent comprises a reagent for measuring the level of the biomarker or the amount thereof by microarray analysis, polymerase chain reaction, reverse transcriptase polymerase chain reaction, Northern blot, Southern blot, or serial analysis of gene expression.
3. The reagent of claim 1, wherein the reagent comprises:
(1) a primer capable of specifically amplifying LINC 02579; or
(2) A probe capable of specifically hybridizing to LINC 02579.
4. The reagent according to claim 3, wherein the primer sequence is shown in SEQ ID No. 5-6.
5. A kit comprising the reagent of any one of claims 1 to 4.
6. A chip comprising the reagent according to any one of claims 1 to 4.
7. Use according to any one of the following:
(1) use of an agent according to any one of claims 1 to 4 in the manufacture of a means for diagnosing allergic airway inflammation;
(2) use of a kit according to claim 5 for the manufacture of a means for diagnosing allergic airway inflammation;
(3) use of the chip of claim 6 for the preparation of a tool for diagnosing allergic airway inflammation.
8. The use according to claim 7, wherein the allergic airway inflammation comprises allergic rhinitis, asthma, allergic rhinitis-asthma syndrome, preferably the allergic airway inflammation is allergic rhinitis-asthma syndrome.
9. A method of screening a candidate drug for the treatment of allergic airway inflammation, comprising the steps of: 1) administering a drug to be tested to a subject in a test group, and detecting the expression level V1 of LINC02579 in a sample derived from said subject in the test group; in a control group, administering a blank control to the subject to be tested, and detecting the expression level V2 of LINC02579 in a sample derived from said subject in the control group; 2) comparing the level V1 and the level V2 detected in the previous step to determine whether the test compound is a candidate for treating allergic airway inflammation.
10. A diagnostic system for performing a computer-implemented method for diagnosing a patient suspected of having allergic airway inflammation, the computer performing steps comprising:
1) accepting input patient data comprising an expression level value for LINC02579 in a biological sample from the patient;
2) analyzing the level of the biomarker and comparing to a corresponding reference value range for the biomarker, if the patient's level of LINC02579 is lower than the reference value range for a non-diseased control subject, indicating that the patient has allergic airway inflammation;
3) displaying diagnostic information about the patient,
wherein the diagnostic system comprises:
1) a storage component for storing data, wherein the storage component has instructions for determining a diagnosis of a subject stored therein;
2) a computer processor for processing data, wherein the computer processor is coupled to the storage component and configured to execute instructions stored in the storage component in order to receive patient data and analyze the patient data according to one or more algorithms; and
3) a display assembly for displaying diagnostic information about the patient;
preferably, the allergic airway inflammation comprises allergic rhinitis, asthma and allergic rhinitis-asthma syndrome, and preferably, the allergic airway inflammation is allergic rhinitis-asthma syndrome.
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