CN106501530A - A kind of biomarker of diagnosing tubercle bacillus infection and its related kit - Google Patents
A kind of biomarker of diagnosing tubercle bacillus infection and its related kit Download PDFInfo
- Publication number
- CN106501530A CN106501530A CN201710007731.2A CN201710007731A CN106501530A CN 106501530 A CN106501530 A CN 106501530A CN 201710007731 A CN201710007731 A CN 201710007731A CN 106501530 A CN106501530 A CN 106501530A
- Authority
- CN
- China
- Prior art keywords
- tuberculosis
- diagnosis
- infection
- cell
- active
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6863—Cytokines, i.e. immune system proteins modifying a biological response such as cell growth proliferation or differentiation, e.g. TNF, CNF, GM-CSF, lymphotoxin, MIF or their receptors
- G01N33/6869—Interleukin
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56911—Bacteria
- G01N33/5695—Mycobacteria
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
- G01N2333/52—Assays involving cytokines
- G01N2333/54—Interleukins [IL]
- G01N2333/5428—IL-10
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
- G01N2333/52—Assays involving cytokines
- G01N2333/54—Interleukins [IL]
- G01N2333/55—IL-2
Abstract
The invention discloses a kind of biomarker of diagnosing tubercle bacillus infection, including IL 2 and IL 10；The present invention confirms that cell factor IL 2 and IL 10 can be used as the marks of tuberculosis infection diagnosis first, can be infected come diagnosis of tuberculosis by the concentration of tuberculosis antigen post-stimulatory IL 2 and IL 10 by determining PMBC, and further discriminate between active tuberculosis and latent tuberculosis infects.The tuberculosis infection blood test reagent prepared based on two kinds of cell factors of the present invention and kit, quick diagnosis for active tuberculosis provide new technical foundation, have Sensitivity and Specificity high, and quick, reasonable is operated, the diagnosis efficiency of active tuberculosis can be significantly improved.
Description
Technical field
The present invention relates to biological diagnosis technical field, more particularly to a kind of infection of diagnosing tubercle bacillus biomarker and
Its related kit.
Background technology
Current immune diagnostic method lungy can not still meet quick early diagnosis tuberculosis and screening latency knot
The needs of core infection.As human body is in the infection of Antituberculous mycobacterium, cell-mediated immune response serves key
Property effect, so finding the T cell antigen of tubercle bacillus specific and detection t cell immune response has become and grinds in recent years
Study carefully focus.
The development and application of IFN-γ release experiment (IFN-γ release assays, IGRAs), to diagnosis lungy
New progress is brought, both experiments also obtain a certain degree of application in clinical detection lungy.IGRAs is applied
The periphery whole blood of tester or detached PMNC (peripheral blood mononuclear cell,
PBMC), after co-culturing with the antigen of tubercle bacillus specific, infected the Memorability in the individual peripheral body of Much's bacillus
T cell can be activated again, so as to promptly discharge including the inflammation factor including IFN-γ.This special by tuberculosis
Property antigenic activation release inflammatory factor such as IIFN- γ, further by MBP enzyme linked immuno-adsorbent assay (enzyme linked
Immunosorbent assay, ELISA) or ELISpot detection (enzyme-linked immunospot assay,
ELISPOT method) is detected, so as to finally judge whether the infection of tubercle bacillus.In recent years, IGRAs is in tuberculosis
It is widely applied in the diagnosis of disease and has been verified, is shown which has in terms of the infection of diagnosis of tuberculosis mycobacterium relatively low
False positive rate and higher sensitiveness, and latent tuberculosis infects can be detected.
Immune response based on detection IFN-γ, has higher sensitiveness in terms of the infection of diagnosing tubercle bacillus, but
Significant deficiency is yet suffered from, its application in diagnosis of tuberculosis is limited, mainly the emission levels of IFN-γ are in activity
No significant difference in tuberculosis patient and latent tuberculosis infects person, therefore current IGRAs be merely able to detect tubercle bacillus sense
Dye, is unable to distinguish active tuberculosis and latent tuberculosis infects, in the regional and national of the high prevalence of tuberculosis, due to latent
The ratio of volt property tuberculosis infection is higher, receives the application in clinical diagnosis of IGRAs and greatly limits.
The existing diagnosis of tuberculosis method based on cell immune response is using IFN-γ as unique diagnosis marker.
IFN-γ is required in host versus protective immunological reaction lungy.But with IFN-γ as unique diagnostic markers
Thing, its sensitiveness yet suffer from deficiency, in addition HIV merge patient lungy or other with immune deficiency tuberculosis sufferer
In person, it may appear that positive control antigen discharge after stimulating IFN-γ amount not enough and cannot result of determination (indeterminate)
Situation, merge in patient lungy in HIV and be more up to 20%-40% therefore, supplement other diagnosis beyond IFN-γ
Mark, can be complementary with IFN-γ improving the sensitiveness of diagnosis.On the other hand due to the difference of the level of release, other lifes
Thing mark combine with IFN-γ be suitable for, can reduce occur cannot result of determination situation.
Additionally, there is research to show, after PMBC is subject to the stimulation of tuberculosis specific antigen, inflammation can be discharged
Mark, the release of these marker of inflammation are considered as related to the immune response of body resisting tuberculosis infection, and ultimately result in
Active tuberculosis and the different outcomes of latent tuberculosis infects.Such as water is discharged by tuberculosis specific antigen post-stimulatory IFN-γ
Flat, constantly can decline with the prolongation for the treatment of time, so as to reflect indirectly the content of molds of body, and IFN-γ burst size increases then
Imply that recurrence lungy.Therefore some marker of inflammation tbas or latent tuberculosis infects institute are peculiar
's.With the development of Luminex suspension chip detection technology, us are made to be possible in the sample of only 25ul while to multiple
Biomarker is detected and is screened.Obtain some cell factors exist in active tuberculosis and latent infection expression poor
Different, the molecular marker of these differential expressions can be as the mark for distinguishing active tuberculosis and latent infection, so as to for tuberculosis
Clinical diagnosis and directiveness treatment provide reference frame.
Content of the invention
The present invention is screened by high-throughput techniques batch for the problem for existing in terms of tubercle bacillus immunodiagnosis at present
And obtain the combination of cytokines that can effectively distinguish active tuberculosis people and latent tuberculosis infects crowd, a kind of tool of exploitation
There are high sensitivity and specificity degree, tuberculosis infection diagnostic reagent kit simple and efficient to handle.
Described cell factor IL-2 and IL-10 are the screenings by high flux suspension chip technology, and obtained can examine
Disconnected mycobacterium tuberculosis infection, and distinguish the new biomarkers of activity and latent tuberculosis infects.
For achieving the above object, the present invention provides a kind of side of the tuberculosis infection biomarker of screening monocyte release
Method, comprises the following steps：
1) preparing different tuberculosis infection state crowds includes active tuberculosis patient, and latent tuberculosis infects person and health are right
PMBC sample according to person
2) using suspension chip technology, the cell factor in sample and chemotactic factor (CF) is detected, obtains cell factor and chemotactic
The data of the factor
3) active tuberculosis patient and the cell factor in latent infection person's sample and Chemokines Levels are compared, by number
According to analysis, the biomarker that can distinguish active tuberculosis and latent tuberculosis infects is obtained
It is yet a further object of the present invention to provide a kind of diagnosing tubercle bacillus infection, and distinguish active tuberculosis and dive
The method of volt property tuberculosis infection.
For achieving the above object, the technical solution used in the present invention is：
Determine what monocyte in the crowd of different tuberculosis infection states was produced after tubercle bacillus specific antigenic stimulus
Cell factor IL-2 and the level of IL-10；
The level of the cell factor for being discharged based on monocyte or being produced, is judged described individual with the presence or absence of tubercle bacillus
Infection.
Further object is that provide one kind infecting for diagnosing tubercle bacillus, and distinguish active tuberculosis
Kit with latent tuberculosis infects.
For achieving the above object, the technical solution used in the present invention is：
A kind of immunodiagnosis kit for diagnosis of tuberculosis mycobacterial infections, the kit energy specific recognition cell
At least one in factor IL-2 and IL-10.
Further, contain containing IL-2 or IL-10 that monocyte produced after stimulation can be determined in mentioned reagent box
The detection components of amount, can be carried out in cell factor IL-2 and IL-10 at least one quantitative.
In a preferred embodiment of the invention, the described detection components to cell factor are ELISA detection groups
Part or electrochemical luminescence detection components.
After the present invention is known, it is possible to use panimmunity correlation technique is detecting containing for IL-2 and IL-10 in sample
Amount, including suspension chip, elispot assay etc., these methods are included in the present invention.
Monocyte used by the present invention may come from detached PMNC and periphery whole blood, can be with
It is cerebrospinal fluid, pleural effusion, ascites, one or more in arthroedema.
Additionally, the present invention relates to mycobacterium, especially Much's bacillus purified protein derivative (purified
Protein derivative, PPD) purposes, for as distinguishing active tuberculosis and latent tuberculosis infects state
Stimulant.
Finally, there is provided containing mycobacterium, the particularly combination of Much's bacillus PPD and ESAT-6 and CFP-10
Thing, detects the tuberculosis infection state of sample in whole blood or monocyte as stimulant
The present invention confirms that cell factor IL-2 and IL-10 can be used as the marks of tuberculosis infection diagnosis first, can be with
Infected come diagnosis of tuberculosis by the concentration of tuberculosis antigen post-stimulatory IL-2 and IL-10 by determining PMBC, gone forward side by side
One step distinguishes active tuberculosis and latent tuberculosis infects.The knot prepared based on two kinds of cell factors of the present invention
Core infects blood test reagent, and the quick diagnosis for active tuberculosis provide new technical foundation, with sensitiveness and specifically
Property high the advantages of, and operate quick, reasonable, the diagnosis efficiency of active tuberculosis can be significantly improved.
Description of the drawings
Fig. 1 is active tuberculosis patient (ATB), latent tuberculosis infects person (LTBI) and normal healthy controls (HC) crowd periphery
Blood monocyte receives the post-stimulatory cytokine release amounts of tuberculosis antigen PPD；
Fig. 2 is the decision-making that joint IL-2 and IL-10 distinguish active tuberculosis (ATB) and latent tuberculosis infects (LTBI)
Tree analysis schematic diagram；
Fig. 3 combines the diagnostic process schematic diagram for active tuberculosis patient suspected for IL-2 and IL-10.
Specific embodiment
The present invention is described in more detail below by specific embodiment, for a better understanding of the present invention,
But following embodiments are not intended to limit the scope of the invention.
Embodiment 1：The screening of the biomarker of diagnosis of tuberculosis mycobacterial infections
Step 1：Case is collected
Collection experimenter's sample, including the blood of active tuberculosis patient, latent tuberculosis infects person and normal healthy controls person
Sample.The foundation that wherein described patient is diagnosed as active tuberculosis is：Patient has Sputum smears positive findings or Sputum culturing positive
As a result, X ray rabat exception or antituberculosis therapy are less than 30 days；The inclusion criteria of latent tuberculosis infects person is：There is tuberculosis sense
The high risk factor of dye has the close contact history of active tuberculosis patient, and tuberculosis is specialOr
Detection is positive, the evidence of rabat and Sputum smears without tuberculosis infection；High risk factor of the normal healthy controls person without tuberculosis infection, without in the recent period
The close contact history of active tuberculosis, rabat or Sputum smears are showed no obvious abnormalities.
According to above-mentioned sample, collection activity tuberculosis patient 25 in the present embodiment, latent tuberculosis infects person 36,
Normal healthy controls person 31.
Step 2：Sample collection and process
Venous blood 5-10ml is adopted extremely from active tuberculosis patient, latent tuberculosis infects person and normal healthy controls person's peripheral blood
In vacuum test tube；
Vacuum test tube is centrifuged, the speed of centrifugation is 2800-3000rpm, and centrifugation time is 10min；It has been centrifuged
Into rear absorption intermediate layer white cell, it is resuspended in cell culture medium RPMI1640, filling-in to 8ml；Upper strata is blood plasma, separable
Frozen standby in -80 DEG C；
4ml Ficoll lymphocyte separation mediums are added in 15ml centrifuge tubes, above-mentioned cell suspension is gently added lymph
Cell separation liquid upper strata, 3000rpm room temperatures are centrifuged 20min；
The intermediate layer cell after centrifugation is drawn in new centrifuge tube, is added RPMI1640 resuspended, is mixed, 2000rpm is centrifuged
5min；
Supernatant is abandoned, 5ml erythrocyte cracked liquids, incubated at room 10min is added；
Add RPMI1640 resuspended to 12ml, mix, 2000rpm is centrifuged 5min, abandons supernatant；Re-suspended cell is in 1ml 1640
In culture medium, cell and typan blue 1 is taken：1 mixing, adds blood cell counting plate to be counted；
In 24 porocyte culture plates, the resuspended PBMC cells of addition 500ml 2.5*10^5/ml；
Tuberculosis specific antigen PPD (10ug/ml) is added per hole, is placed into after mixing in 37 DEG C of cell culture incubators, cultivated
18-24 hours；’
Nutrient solution supernatant is sucked, 200-300ul can be drawn per hole, be transferred in 1.5ml EP pipes；
Step 3：Suspension chip technology is detected
The core technology of suspension chip is small particle or to claim microballoon (bead) to dye different iridescent respectively, so
Again the albumen or oligonucleotide probe for different testing sample is adsorbed onto on the microballoon of different colours in the way of covalent afterwards.Micro-
Ball mixes two kinds of different red classification fluorescent dyes according to strict blend proportion in manufacturing process, can according to ratio difference
So that sphere matrix is categorized as 100 kinds, every kind of put on different probe molecules, such that it is able to up to 100 kinds in a sample
Different molecules of interest synchronizes detection.
This research is to active tuberculosis, three groups of crowd's Peripheral Bloods of latent tuberculosis infects and normal healthy controls and specificity
Cell culture supernatant after antigenic stimulus detected, its totally 27 kinds of cell factor for detecting：IL-1beta,IL-1ra,IL-
2,IL-4,IL-5,IL-6,IL-7,IL-8,IL-9,IL-10,IL-12(p70),IL-13,IL-15,IL-17,basic FGF,
eotaxin,G-CSF,GM-CSF,IFN-γ,IP-10,MCP-1(MCAF),MIP-1alpha,MIP-1beta,PDGF-BB,
RANTES,TNF-alpha,VEGF
Bio-Plex suspension chip operations are comprised the following steps that：
Prepare standard items, standard items are resuspended with 500ul sample diluents, 30mim is incubated on ice after concussion 1-3s, according to
The requirement dilution standard product of specification；
All standard items, sample are placed room temperature 20min balance, quickly 10 × magnetic bead is centrifuged, 5ul 10 is needed per hole
1 × magnetic bead middling speed whirlpool is shaken 30s by × magnetic bead, adds 50ul per hole；
Standard items are gently shaken, is added after control and sample 1-3s in hole, per hole 50ul.Incubated at room 30min.Centrifugation with
Supernatant discarded afterwards；
Detection antibody is added in per hole, is cultivated 30 minutes, washed 3 times；
Streptavidin-PE is added, after the completion of 30min incubations, then is washed 3 times, in every hole, add nitrite ion, room temperature is incubated
Educate 10min.After the completion of wash again 3 times addition buffer solutions；
Sample to be tested further carries out reading and detection in Luminex systems, and applies corresponding analysis software to carry out
Analysis.The concentration application pg/ml of final inflammatory factor is calculated as 0pg/ml less than the concentration value of Monitoring lower-cut representing.
Step 4：Screening biomarker simultaneously sets up forecast model
According to content of the 27 kinds of cell factors for being obtained in different crowd, we are therefrom filtered out in active tuberculosis
There is the cell factor of differential expression in people and latent tuberculosis infects person：The post-stimulatory IL-2 of PPD, IL-10, IP-10, IFN-
There is notable difference in the expression of γ and TNF-α, can infect the biological marker of different conditions between the two groups as diagnosis of tuberculosis
Thing (Fig. 1).In order to further analyze the diagnostic value of these cell factors, we have carried out experimenter's operating curve first
The analysis of (Receiver-operating-characteristic, ROC), is used with the TG-AUC (AUCs) in ROC analyses
To evaluate individual gene to two groups of the separating capacity of being located.AUCs values are bigger, illustrate that the gene pairs two groups of the separating capacity is got over
Good.
Mould is predicted to further set up for distinguishing the cell factor of active tuberculosis and latent tuberculosis infects
Type, we employ decision tree modeling, and the software of employing is R program software.The best of breed base of molecular marker
In cross validation and minimal error principle, the nodal values at minimum branch node are 10.By decision tree analysis, cell is obtained
The best of breed of the factor be IL-2 and IL-10, both united diagnostic sensitivities and specificity be respectively 84.0% and 88.9%,
The accuracy of diagnostic result is 86.9% (Fig. 2).
The present embodiment filters out between active tuberculosis patient and latent tuberculosis infects person difference table in PBMC cells
The cell factor for reaching.With the cell factor that filtered out as diagnosis marker, application suspension chip technology or elisa technique are carried out
Detection, can be used in the quick detection of active tuberculosis blood, with higher sensitivity height and the degree of accuracy, simple easily behaviour
Make, the diagnosis efficiency of active tuberculosis can be greatly improved.
Embodiment 2：Application of the biomarker and its related kit of diagnosing tubercle bacillus infection in clinic
Step 1：Case is collected
Clinically collection activity patient suspected lungy, inclusion criteria is：There are similar active tuberculosis
Clinical manifestation is such as coughed, spits blood, becoming thin, generating heat and night sweat etc., or shows with the impact similar with active tuberculosis,
There is the high risk factor of tuberculosis infection or the close contact history of active tuberculosis patient.In this example, according to above-mentioned selected mark
Standard, collects doubtful active tuberculosis patient 44 altogether.
Step 2：Specimen collection and process
Venous blood 5-10ml is adopted from the peripheral blood of collected active tuberculosis patient suspected into vacuum test tube；Enter
One step separating peripheral blood mononuclear cells：Vacuum test tube is centrifuged, the speed of centrifugation is 2800-3000rpm, during centrifugation
Between be 10min；Intermediate layer white cell is drawn after the completion of centrifugation, is resuspended in cell culture medium RPMI1640, is added 4ml
Above-mentioned cell suspension is gently added lymphocyte separation medium upper strata, 3000rpm room temperatures to be centrifuged by Ficoll lymphocyte separation mediums
20min；The intermediate layer cell after centrifugation is drawn in new centrifuge tube, is added RPMI1640 resuspended, is mixed, 2000rpm is centrifuged
5min；Supernatant is abandoned, 5ml erythrocyte cracked liquids, incubated at room 10min is added；Add RPMI1640 resuspended to 12ml, mix,
2000rpm is centrifuged 5min, abandons supernatant；Re-suspended cell takes cell and typan blue 1 in 1640 culture mediums of 1ml：1 mixing,
Blood cell counting plate is added to be counted；
The PBMC for having counted, for carrying out T-SPOT detections, another part is added in 24 porocyte culture plates a part, per
Hole adds tuberculosis specific antigen PPD (10ug/ml), is placed in 37 DEG C of cell culture incubators after mixing, cultivates 18-24 hours；
Nutrient solution supernatant is sucked, is transferred in 1.5ml EP pipes, for further detection.
Step 3, the content that IL-2 and IL-10 is detected using elisa technique：
The content application ELISA method of L-2 and IL-10 is measured.To apply Duoset ELISA Development
As a example by kit (R＆D Systems Inc, MN, USA) is detected, operating procedure is carried out fully according to operating instruction, briefly
Step is as follows：1). 96 holes of application are flat ELISA Plate (NUNC maxisorp, Roskilde, Denmark), are then coated per hole
Capture antibody (capture antibody) after 100ul dilutions, 4oC are coated overnight；2) coating overnight washes away coating buffer afterwards, should
At least 1h is closed with containing 1% bovine serum albumin(BSA) (bovine serum albumin, BSA)；3) cleaning solution washes 3 times
Afterwards, the sample and corresponding standard items after adding 100ul suitably to dilute per hole, is incubated 2h under room temperature；4) after washing 3 times, per
Hole adds detection antibody, incubated at room 2h；5) after washing 3 times, the addition Streptavidin-HRP per hole, incubated at room 20min,
Nitrite ion TMB is added after washing 3 times, after lucifuge is reacted to time enough, terminate liquid 2N H2SO4 terminating reactions is applied；6) should
The absorbance OD values per hole in 450nm are read with ELIASA, and applies the correction of 540nm；7) according to the OD values of standard items and
Corresponding concentration makes calibration curve, then calculates the concentration of sample according to calibration curve, is multiplied by extension rate and for actual concentrations,
The thinner ratio of IL-2 and IL-10 is 1 in our current research:2；8) all samples of this research are all multiple holes, ultimate density application pg/
Ml is represented
Step 4, the judgement of diagnostic result：
The diagnositc decision method based on two kinds of marks of IL-2 and IL-10 is specifically provided in the embodiment of the present invention, specifically
Method can be found in Fig. 2：
Burst size such as IL-2 is more than or equal to 282.5pg/ml, while the burst size of IL-10 is more than or equal to 113.9pg/ml,
Testing result is the positive, that is, be judged as active tuberculosis patient；If the burst size of IL-2 is less than 282.5pg/ml, or IL-10
Burst size be less than 113.9pg/ml, testing result for feminine gender, that is, be judged as the patient of latent tuberculosis infects.In this example
In 61 samples, used in (25 active tuberculosis patients, 36 latent tuberculosis infects persons), the diagnosis of the decision method is sensitive
Property up to 84.0%, up to 88.9%, the accuracy of differentiation is 86.9% (Fig. 3) to specificity.
It is able to know that by above-described embodiment, the present invention confirms that cell factor IL-2 and IL-10 can be used as tuberculosis first
The mark of Infect And Diagnose, can pass through to determine PMBC by the dense of tuberculosis antigen post-stimulatory IL-2 and IL-10
Degree carrys out diagnosis of tuberculosis infection, and further discriminates between active tuberculosis and latent tuberculosis infects.With two kinds of the present invention
The tuberculosis infection blood test reagent prepared based on cell factor, the quick diagnosis for active tuberculosis provide new technology
Basis, high have the advantages that Sensitivity and Specificity, and quick, reasonable is operated, active tuberculosis can be significantly improved
Diagnosis efficiency.
Above the specific embodiment of the present invention is described in detail, but which has been intended only as example, the present invention has not been limited
It is formed on particular embodiments described above.To those skilled in the art, any equivalent modifications carried out by the present invention and
Replacement is also all among scope of the invention.Therefore, the impartial conversion made without departing from the spirit and scope of the invention and
Modification, should all cover within the scope of the invention.
Claims (2)
1. the biomarker that a kind of diagnosing tubercle bacillus infect, it is characterised in that the biomarker is IL-2 and IL-
10.
2. a kind of for diagnosing tubercle bacillus infection kit, it is characterised in that include IL-2 in the kit special
Property antibody and/or IL-10 specific antibodies.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710007731.2A CN106501530A (en) | 2017-01-05 | 2017-01-05 | A kind of biomarker of diagnosing tubercle bacillus infection and its related kit |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710007731.2A CN106501530A (en) | 2017-01-05 | 2017-01-05 | A kind of biomarker of diagnosing tubercle bacillus infection and its related kit |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106501530A true CN106501530A (en) | 2017-03-15 |
Family
ID=58345021
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710007731.2A Pending CN106501530A (en) | 2017-01-05 | 2017-01-05 | A kind of biomarker of diagnosing tubercle bacillus infection and its related kit |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106501530A (en) |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107478563A (en) * | 2017-07-19 | 2017-12-15 | 浙江省人民医院 | A kind of method based on the double cell factors of Flow cytometry tuberculosis specificity |
| CN107632155A (en) * | 2017-09-06 | 2018-01-26 | 上海兰卫医学检验所股份有限公司 | A kind of blood cell is to the immunoreactive detection method of tubercle bacillus specific |
| CN107653313A (en) * | 2017-09-12 | 2018-02-02 | 首都医科大学附属北京胸科医院 | Applications of the RETN and KLK1 in as diagnosis mark |
| CN108226535A (en) * | 2018-01-19 | 2018-06-29 | 中国人民解放军第三〇九医院 | Application of the system of detection adhesion molecule and cytokine content in retreat tuberculosis patient outcomes are detected |
| CN108845129A (en) * | 2018-06-01 | 2018-11-20 | 广东医科大学 | A kind of application of the biomarker of active tuberculosis class disease |
| CN113884685A (en) * | 2021-10-18 | 2022-01-04 | 中国农业大学 | Bovine tuberculosis serological diagnosis marker and clinical application thereof |
| CN114859041A (en) * | 2021-02-03 | 2022-08-05 | 首都医科大学附属北京胸科医院 | Detection method and reagent for tubercle bacillus |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7838013B2 (en) * | 2003-05-08 | 2010-11-23 | Statens Serum Institut | Specific epitope based immunological diagnosis of tuberculosis |
| WO2012167307A1 (en) * | 2011-06-06 | 2012-12-13 | The University Of Melbourne | Diagnosis of mycobacterial infection |
| CN104541169A (en) * | 2012-07-16 | 2015-04-22 | 捷狮有限公司 | Status of tuberculosis infection in an individual |
| CN105218648A (en) * | 2015-07-10 | 2016-01-06 | 邓林红 | Negre antigen, nucleotide sequence and application thereof |
| CN105384805A (en) * | 2015-10-16 | 2016-03-09 | 上海市肺科医院 | Tuberculosis detection polypeptide and application thereof |
| CN105861520A (en) * | 2015-01-20 | 2016-08-17 | 广东体必康生物科技有限公司 | Application of Rv3121 protein for detecting mycobacterium tuberculosis infection |
-
2017
- 2017-01-05 CN CN201710007731.2A patent/CN106501530A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7838013B2 (en) * | 2003-05-08 | 2010-11-23 | Statens Serum Institut | Specific epitope based immunological diagnosis of tuberculosis |
| WO2012167307A1 (en) * | 2011-06-06 | 2012-12-13 | The University Of Melbourne | Diagnosis of mycobacterial infection |
| CN104541169A (en) * | 2012-07-16 | 2015-04-22 | 捷狮有限公司 | Status of tuberculosis infection in an individual |
| CN105861520A (en) * | 2015-01-20 | 2016-08-17 | 广东体必康生物科技有限公司 | Application of Rv3121 protein for detecting mycobacterium tuberculosis infection |
| CN105218648A (en) * | 2015-07-10 | 2016-01-06 | 邓林红 | Negre antigen, nucleotide sequence and application thereof |
| CN105384805A (en) * | 2015-10-16 | 2016-03-09 | 上海市肺科医院 | Tuberculosis detection polypeptide and application thereof |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107478563A (en) * | 2017-07-19 | 2017-12-15 | 浙江省人民医院 | A kind of method based on the double cell factors of Flow cytometry tuberculosis specificity |
| CN107632155A (en) * | 2017-09-06 | 2018-01-26 | 上海兰卫医学检验所股份有限公司 | A kind of blood cell is to the immunoreactive detection method of tubercle bacillus specific |
| CN107653313A (en) * | 2017-09-12 | 2018-02-02 | 首都医科大学附属北京胸科医院 | Applications of the RETN and KLK1 in as diagnosis mark |
| CN107653313B (en) * | 2017-09-12 | 2021-07-09 | 首都医科大学附属北京胸科医院 | Application of RETN and KLK1 as tuberculosis detection markers |
| CN108226535A (en) * | 2018-01-19 | 2018-06-29 | 中国人民解放军第三〇九医院 | Application of the system of detection adhesion molecule and cytokine content in retreat tuberculosis patient outcomes are detected |
| CN108845129A (en) * | 2018-06-01 | 2018-11-20 | 广东医科大学 | A kind of application of the biomarker of active tuberculosis class disease |
| CN108845129B (en) * | 2018-06-01 | 2021-03-30 | 广东医科大学 | Application of biomarker of active tuberculosis diseases |
| CN114859041A (en) * | 2021-02-03 | 2022-08-05 | 首都医科大学附属北京胸科医院 | Detection method and reagent for tubercle bacillus |
| CN113884685A (en) * | 2021-10-18 | 2022-01-04 | 中国农业大学 | Bovine tuberculosis serological diagnosis marker and clinical application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106501530A (en) | A kind of biomarker of diagnosing tubercle bacillus infection and its related kit | |
| Yao et al. | Multiplex analysis of plasma cytokines/chemokines showing different immune responses in active TB patients, latent TB infection and healthy participants | |
| CN108802385A (en) | Label and determinant for diagnosing infection and its application method | |
| CN104020297B (en) | Test kit for m tuberculosis infection detection and clinical therapeutic efficacy monitoring and application thereof | |
| CN104823052A (en) | Biomarkers for diagnosing and/or monitoring tuberculosis | |
| WO2018076404A1 (en) | Method for in vitro detection of active tuberculosis | |
| EP2791679A1 (en) | Methods and kits for diagnosing latent tuberculosis infection | |
| CN103760345B (en) | A kind of kit and application thereof utilizing peripheral blood detection m tuberculosis infection | |
| CN103954754A (en) | Immunologic diagnosis kit of active tuberculosis | |
| CN103063836B (en) | Detect the reagent of mycobacterial infections, method and kit | |
| CN106939035A (en) | A kind of mycobacterium tuberculosis T cell antigen epitope polypeptide and its application | |
| CN103063837B (en) | Reagent, method and kit for detecting mycobacterial infection | |
| CN109991417B (en) | Immune marker for tuberculosis and application | |
| KR101733864B1 (en) | Biomarker for differentiating between active and latent tuberculosis in conjunction with Interferon gamma release assay and its use | |
| CN114675036A (en) | Marker for diagnosing T-SPOT negative tuberculosis and application thereof | |
| CN109813903A (en) | The method for being used for diagnosis of tuberculosis mycobacterial infections based on Flow cytometry tuberculosis Specific T cell immunity phenotype | |
| RU2756314C2 (en) | Tuberculosis biomarkers | |
| CN107860907A (en) | Application of the albumen of Pru p 3 in wormwood artemisia pollen correlation peach allergy detection kit is prepared | |
| Miyata et al. | Cellular immunity reflects the persistent symptoms among COVID-19 recovered patients in Japan | |
| CN107976543A (en) | A kind of diagnosis kit and detection method | |
| CN117451989A (en) | Immune marker capable of predicting prognosis curative effect of allergic rhinitis immunotherapy and application | |
| CN117269491A (en) | Biomarker capable of predicting prognosis curative effect of allergic rhinitis immunotherapy and application | |
| CN103675293A (en) | Application of proteins of Rv3872, Rv0164 and/or Rv1926c in developing and/or designing product with functions of identification, diagnosis, auxiliary diagnosis, screening and/or auxiliary screening of active tuberculosis | |
| CN103525829A (en) | Preparation method of recombinant antigen of mycobacterium tuberculosis capable of being used for diagnosing tuberculosis infection | |
| KR102368216B1 (en) | Process for detecting latent infected cow with bovine tuberculosis using ELISA boosting method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20170315 |