CN106546737A - A kind of method of vitro detection active tuberculosis - Google Patents
A kind of method of vitro detection active tuberculosis Download PDFInfo
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- CN106546737A CN106546737A CN201610925713.8A CN201610925713A CN106546737A CN 106546737 A CN106546737 A CN 106546737A CN 201610925713 A CN201610925713 A CN 201610925713A CN 106546737 A CN106546737 A CN 106546737A
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Abstract
The invention discloses a kind of method of vitro detection active tuberculosis.Methods described is to detect the application in tuberculosis in vitro based on the reagent of quantitative determination tuberculosis correlation factor IFN γ and IL 2.Compared with existing active tuberculosis detection method, this method is more accurate in the judged result of active tuberculosis, specificity is higher.Compared with existing goldstandard Sputum culturing, detection time is greatly shortened, and is diagnosed patient, direction of medication usage in time for clinician and is provided foundation, to control lungy with very positive meaning.It is including pulmonary tuberculosis, scrofula, intestinal tuberculosis, bone tuberculosis, nephrophthisis etc. for all tuberculosis that heretofore described reagent is used for the tuberculosis of detection, and coverage rate and practicality are wider.
Description
Technical field
The invention belongs to detection field, and in particular to a kind of method of vitro detection active tuberculosis.
Background technology
Tuberculosis is a kind of common chronic infectious disease caused by Much's bacillus, can be invaded and many internal organs, with lung
Portion's tuberculosis infection is most commonly seen.The whole world has nearly 1/3 people to infect tubercle bacillus at present, and statistics shows have in 2013
1500000 people die from tuberculosis, 9,000,000 new cases, and tuberculosis has become the main disease of whole world adult's death because of infectious disease
One of disease.China is one of high burden country of 22, whole world tuberculosis, and active tuberculosis patient's number occupies second place of the world.Tuberculosis
The infection of disease mainly occurs undiscovered and before being treated in patient, and 1 tuberculosis patient is in 1 year by connecing closely
Touch and can infect up to 10-15 people.Circulation way lungy is Jing respiratory infectious, in the lung of pulmonary tuberculosis patient has substantial amounts of
Tubercle bacillus, just probably by pathogen transmission to Healthy People if he is coughed or sneezed against healthy population.And
It was found that before sufferer, patient does not take any preventive means, during with the close contact such as kinsfolk, colleague, classmate,
Contactee is easy for by tubercle bacillus affection.Primary treatments master lungy is drug therapy at this stage, but such is controlled
Treatment can not prevent propagation lungy completely.Bcg vaccination (BCG) is current internationally recognized most effective prevention tuberculosis
Immunization method, China planned immunization program in specify neonate must bcg vaccination, but its protective efficacy is also simultaneously
Not up to 100%, therefore, prevent primary measure lungy to be the neopathy people for finding as early as possible to be hidden in crowd.
Clinic is usually used in the method for active tuberculosis diagnosis at present, Imaging Method poor specificity, diagnoses process with doctor
Raw experience is closely related, and diagnostic result carries stronger subjectivity；Sputum culturing method is Diagnosis of Tuberculosis goldstandard, but sensitiveness
It is low, and time-consuming (2-8 is all), is unfavorable for the early diagnosis of tuberculosis；Ziehi-Neelsen stain sensitiveness only has 20-30%；Molecular diagnosis
Method (such as PCR) poor repeatability, and it is cumbersome, there is certain requirement to laboratory environment；Tuberculin test (TST) is easily received
The impact that BCG vaccine (BCG) is inoculated with, causes the false positive of testing result.γ-the interference based on T cell is occurred in that in recent years
Plain release test, but the method is only capable of judging whether patient has infected (mistake) Mycobacterium tuberculosis, it is impossible to determine whether to live
Dynamic property tuberculosis.In above detection method, although Sputum culturing, Ziehi-Neelsen stain and molecular diagnosis method specificity is good, only for
Bacterium yang constipation core (accounting for 30%) in tuberculosis patient and bacterium yin constipation core can not be diagnosed, clinically still lack at present it is a kind of directly
Method active tuberculosis is detected.
Much's bacillus is a kind of typical intracellular infection bacterium, and human body is to its immunologic mechanism mainly based on T cell
Cellular immunity.After Much's bacillus invades human body, when immunity of organisms is low, macrophage can not prevent from being gulped down
The M. tuberculosis growth bitten, but the Much's bacillus of phagocytosis can be taken to elsewhere, offer antigen, now tuberculosis branch bar
Bacterium amount reproduction, makes the T lymphocytes of T lymphocytes sensitization around, sensitization produce cytokine profiles (such as INF- γ, IL-
2 and TNF-α etc.), these cell factor collective effects kill the Much's bacillus in focus.
The content of the invention
In order to solve above-mentioned problem, the present invention is had found anti-through differential stimulus by substantial amounts of screening experiment
In cell factor secreted by the PMNC of former induction, IFN-γ and IL-2 are had with the morbid state of patient closely
Contact, by detecting whether the secretory volume of the two factors can be that active tuberculosis is accurately judged to patient, more favorably
Diagnosis is made in time to the morbid state of patient in clinician.
The technical solution used in the present invention is：
The reagent of quantitative determination tuberculosis correlation factor IFN-γ and IL-2 detects the application in tuberculosis in vitro.
Preferably, quantitative detecting reagent includes enzyme linked immunosorbent assay, enzyme-linked immunospot assay, chemoluminescence method, liquid
Reagent used by chip method.
Preferably, tuberculosis is active tuberculosis.
Preferably, active tuberculosis include pulmonary tuberculosis, activity scrofula, activity intestinal tuberculosis, activity bone tuberculosis,
Activity nephrophthisis etc..
A kind of method of vitro detection active tuberculosis, it is thin using the single core of differential stimulus antigen in vitro inducing peripheral blood
Born of the same parents so as to produce tuberculosis correlation factor, by the secretory volume of detection wherein IFN-γ and IL-2, whether judgement sample is activity
Tuberculosis, the method are not used in medical diagnosis on disease.
Preferably, specific antigen includes at least one of tri- kinds of albumen of ESAT-6, CFP-10 and Rv1985c, form choosing
Fusion protein, protein mixture or peptide fragment from these three albumen.
Preferably, detect that method used includes enzyme linked immunosorbent assay, enzyme-linked immunospot assay, chemoluminescence method, liquid
State chip method.
The invention has the beneficial effects as follows：
1) the invention provides a kind of effective method for judging active tuberculosis, the method is by stimulating PMNC
Tuberculosis relevant cell factor is produced, the detection wherein secretory volume of IFN-γ and IL-2 is simultaneously analyzed, it was therefore concluded that.With it is existing
Active tuberculosis detection method is compared, and this method is more accurate in the judged result of active tuberculosis, specificity is higher.
2), compared with existing goldstandard Sputum culturing, detection time is significantly for the active tuberculosis detection method that the present invention is provided
Shorten, patient, direction of medication usage are diagnosed in time for clinician and provide foundation, to control lungy with very positive meaning
Justice.
3) heretofore described reagent be used for detection tuberculosis be for all tuberculosis, including outside pulmonary tuberculosis and lung tie
Core (such as scrofula, intestinal tuberculosis, bone tuberculosis, nephrophthisis etc.), coverage rate and practicality it is wider.
Description of the drawings
Fig. 1 is the secretion level that ELISA detects IFN-γ and IL-2；
Fig. 2 is the secretion level that ELISPOT detects IFN-γ and IL-2；
Fig. 3 is ELISA detection IFN-γs, the secretion level of IL-2 and IL-10.
Specific embodiment
Active tuberculosis：Refer to tuberculosis, it was demonstrated that have Much's bacillus to discharge, focus belongs to active stage, rabat
On often have patch shape shade or tuberculous cavity, or send out focus, illustrate that Much's bacillus breeding is active, virulence is strong.It is living
Dynamic property tubercular is major source of infection lungy, such as can not diagnoses and treatment in time, can cause it is lungy propagate on a large scale,
It is the key for reducing incidence of tuberculosis and case fatality rate fast and accurately to detect active tuberculosis patient.
Active tuberculosis includes but is not limited to active tuberculosis, activity scrofula, activity intestinal tuberculosis, activity
Bone tuberculosis, activity nephrophthisis.
With reference to specific embodiment, the present invention is further illustrated, but is not limited thereto.
Embodiment 1：Density-gradient centrifugation method separates human peripheral blood single nucleus cell (PBMC)
It is using the heparin tube aseptic aspiration personnel to be tested peripheric venous blood 4-5ml of fresh heparin lithium or liquaemin, reverse to make anti-freezing
Agent and blood mix stand-by.Prepare 2 15ml centrifuge tubes, be separately added into and the isopyknic injection physiological saline of blood sampling volume and pouring
Bar cell separation liquid.After fresh heparin anti-coagulating is mixed with physiological saline, uniform speed slow is added in separating liquid, is needed during addition
Separating liquid is kept to be layered with blood, then in 22 DEG C, 1800g centrifugation 20min, visible PBMCs cells are present in cloud and mist after centrifugation
In shape layer.PBMCs cellular layers are drawn to new 15ml centrifuge tubes, with RPMI-1640 nutrient solutions polishing to 12ml, in 22 DEG C,
600g is centrifuged 10min.Supernatant is outwelled, with RPMI-1640 nutrient solutions polishing to 5ml, after gently piping and druming precipitation is resuspended, in 22 DEG C,
350g is centrifuged 10min.Supernatant is abandoned, 0.5ml serum free medium re-suspended cells are added.Using Trypan Blue manual count method or
Automatically counted using suitable equipment.According to count results, the PBMC for being diluted to desired concn with serum free medium is thin
Born of the same parents' suspension.
2 enzyme linked immunosorbent assay of embodiment (ELISA) detects the induction of clinical diagnosis active tuberculosis patient and Healthy People
The level of PBMC secretion of gamma-IFN and IL-2 afterwards
1) choose the liquaemin anti-freezing peripheral vein blood sample of patient and 10 Healthy Peoples of 10 clinical diagnosises for active tuberculosis
This, is separated PBMC by the density stratification method in embodiment 1, is diluted to PBMC using serum free medium after cell count
2.5×106The concentration of individual/ml, cell is seeded in 96 orifice plates, and 2.5 × 10 are added per hole5Individual cell, adds and uses serum-free
The differential stimulus antigen (fusion protein CFP-10-ESAT-6-Rv1985c, 2 μ g/ml of concentration) of culture medium dilution, 37 DEG C, 5%
CO2Under the conditions of be incubated 18 hours, taking cell culture fluid carries out the detection of IFN-γ and IL-2 cell factors.
2) using the elisa plate for being coated with anti-human IFN-γ monoclonal antibody in advance, walk in carried cell culture in addition
Clear liquid, is incubated at room temperature 2 hours, clean elisa plate 3-5 time with lavation buffer solution, adds anti-IFN-γ antibody, room toward detecting in hole
Temperature incubation 1 hour, then removes antibody, clean elisa plate 3-5 time with lavation buffer solution, and add that HRP marks two are anti-, and 37
DEG C incubation 1 hour, then removes two and resists, and clean elisa plate 3-5 time with lavation buffer solution, and the TMB that addition is prepared develops the color bottom
Thing, under room temperature, lucifuge develops the color 20 minutes, is eventually adding terminate liquid color development stopping, carries out interpretation of result with ELIASA in 30 minutes.
3) using the elisa plate for being coated with anti-human IL-2 monoclonal antibodies in advance, in addition, walk put forward cells and supernatant
Liquid, is incubated at room temperature 2 hours, clean elisa plate 3-5 time with lavation buffer solution, adds anti-IL-2 antibody, room temperature to incubate toward detecting in hole
Educate 1 hour, then remove antibody, elisa plate 3-5 time is cleaned with lavation buffer solution, add HRP marks two resist, and 37 DEG C incubate
Educate 1 hour, then remove two and resist, clean elisa plate 3-5 time, the TMB chromogenic substrates that addition is prepared, room with lavation buffer solution
The lower lucifuge of temperature develops the color 20 minutes, is eventually adding terminate liquid color development stopping, carries out interpretation of result with ELIASA in 30 minutes.
Testing result is as shown in figure 1, it can be seen that compared with Healthy People, the PBMC of active tuberculosis patient is being passed through
After differential stimulus antigen induction, the secretory volume of its IFN-γ and IL-2 both cell factors is all significantly improved.With IFN-γ
Secretory volume ＞ 20, IL-2 secretory volumes ＞ 15 are standard, and the sample that two cytokines measurement results are positive is can determine whether as work
Dynamic property tuberculosis.According to this decision scheme, the 10 active tuberculosis clinical samples detected with the method are all judged to activity
Tuberculosis, 10 Healthy People sample standard deviations are judged to inactive tuberculosis.
3 enzyme-linked immunospot assay of embodiment (ELISPOT) detects luring for clinical diagnosis active tuberculosis patient and Healthy People
Lead the level of rear PBMC secretion of gamma-IFN and IL-2
1) the liquaemin anti-freezing peripheral vein blood sample of 10 active tuberculosis patients and 10 Healthy Peoples is chosen, by embodiment 1
In density stratification method separate PBMC, be diluted to 2.5 × 10 using serum free medium after cell count6Individual/ml
Concentration it is standby.
2) pvdf membrane detection plate is selected (to use anti-human IFN-γ monoclonal antibody and anti-human IL-2 monoclonals anti-respectively in advance
Body is coated with overnight, and 4 DEG C of conditions are preserved) as reaction plate, 2.5 × 10 are added per hole5PBMC prepared by individual upper step, each sample are every
Kind of cell factor arranges 3 detection holes, is that blank (addition serum free medium), experimental port (add specificity thorn respectively
Sharp antigen is stimulated, concentration be 2 μ g/ml) and Positive control wells (add phytolectin PHA stimulated, concentration be 5 μ g/
Ml), 37 DEG C, 5%CO216-20 hours are incubated in incubator.
3) detection plate is taken out, abandon culture supernatant, detection plate 3-4 time is rinsed with PBS, then with PBS board-washings 1-2 time, every time
Gently rocked under 5 back and forth, patted dry after last time board-washing.In each hole, add the labelled antibody of corresponding cell factor to work
Liquid, puts 37 DEG C and is incubated 1 hour.Liquid is abandoned, with PBS board-washings 3-5 time, is gently rocked back and forth under 5 every time, clapped after last time board-washing
It is dry, enzyme conjugates working solution is added per hole, 37 DEG C is put and is incubated 1 hour.Liquid is abandoned, with PBS board-washings 5 times, is gently shaken back and forth every time
Under dynamic 5, pat dry after last time board-washing, the nitrite ion of 100 μ l is added per hole, put 37 DEG C of lucifuge colour developing 5-10 minutes big to spot
Little naked eyes are high-visible, abandon liquid, deionized water or originally water washing detection plate positive and negative and base 3-5 time, in button dry hole
Water, plate is put into 37 DEG C of oven dryings 4 hours or dry overnight at room temperature by color development stopping.Colour developing result CTL
ImmunoSpot S6 ELISpot analyzers are automatically analyzed.
Experimental analysis is tied as shown in Fig. 2 it can be seen that the PBMC of active tuberculosis patient is through differential stimulus
After antigen induction, the secretory volume of its IFN-γ and IL-2 is all significantly improved.
Embodiment 4 carries out induction stimulation to the PBMC of active tuberculosis patient, latent infection and Healthy People respectively, and to which
The secretory volume of middle IFN-γ, IL-2 and IL-10 is contrasted
1) choose the liquaemin anti-freezing peripheral vein blood sample of 10 active tuberculosis patients, 10 latent infections and 10 Healthy Peoples
This, is separated PBMC by the density stratification method in embodiment 1, is diluted to PBMC using serum free medium after cell count
2.5×106The concentration of individual/ml, cell is seeded in 96 orifice plates, and 2.5 × 10 are added per hole5Individual cell, adds and uses serum-free
The differential stimulus antigen (2 μ g/ml of concentration) of culture medium dilution, 37 DEG C, 5%CO2Under the conditions of be incubated 18 hours, take cell culture
Liquid carries out the detection of IFN-γ, IL-2 and TNF-α cell factor.
2) using the elisa plate for being coated with anti-human IFN-γ monoclonal antibody in advance, walk in carried cell culture in addition
Clear liquid, is incubated at room temperature 2 hours, clean elisa plate 3-5 time with lavation buffer solution, adds anti-IFN-γ antibody, room toward detecting in hole
Temperature incubation 1 hour, then removes antibody, clean elisa plate 3-5 time with lavation buffer solution, and add that HRP marks two are anti-, and 37
DEG C incubation 1 hour, then removes two and resists, and clean elisa plate 3-5 time with lavation buffer solution, and the TMB that addition is prepared develops the color bottom
Thing, under room temperature, lucifuge develops the color 20 minutes, is eventually adding terminate liquid color development stopping, carries out interpretation of result with ELIASA in 30 minutes.
3) using the elisa plate for being coated with anti-human IL-2 monoclonal antibodies in advance, in addition, walk put forward cells and supernatant
Liquid, is incubated at room temperature 2 hours, clean elisa plate 3-5 time with lavation buffer solution, adds anti-IL-2 antibody, room temperature to incubate toward detecting in hole
Educate 1 hour, then remove antibody, elisa plate 3-5 time is cleaned with lavation buffer solution, add HRP marks two resist, and 37 DEG C incubate
Educate 1 hour, then remove two and resist, clean elisa plate 3-5 time, the TMB chromogenic substrates that addition is prepared, room with lavation buffer solution
The lower lucifuge of temperature develops the color 20 minutes, is eventually adding terminate liquid color development stopping, carries out interpretation of result with ELIASA in 30 minutes.
4) using the elisa plate for being coated with anti-human IL-10 monoclonal antibodies in advance, in addition, walk put forward cells and supernatant
Liquid, is incubated at room temperature 2 hours, clean elisa plate 3-5 time with lavation buffer solution, adds anti-IL-2 antibody, room temperature to incubate toward detecting in hole
Educate 1 hour, then remove antibody, elisa plate 3-5 time is cleaned with lavation buffer solution, add HRP marks two resist, and 37 DEG C incubate
Educate 1 hour, then remove two and resist, clean elisa plate 3-5 time, the TMB chromogenic substrates that addition is prepared, room with lavation buffer solution
The lower lucifuge of temperature develops the color 20 minutes, is eventually adding terminate liquid color development stopping, carries out interpretation of result with ELIASA in 30 minutes.
Experimental result is as shown in figure 3, according to IFN-γ secretion amount ＞ 20, IL-2 secretory volumes ＞ 15 and IL-10 secretory volumes ＞
25 respectively as the positive criterion of three cytokines measurements：
1), in 10 active tuberculosis samples, 9 are judged as that IFN-γ is positive, and 9 are judged as that IL-2 is positive, and 4 are judged as IL-
10 is positive, and the cross sample collection that these three factors judge is as shown in Table 1.
Table 1
According to table 1, as can be seen that IFN-γ and IL-2 the two factors to be combined, can determine that 8 samples be positive, and be increased
The IL-10 factors are judged that the sensitivity to improving detection is not helped.
2), in 10 latent infection samples, 6 are judged as that IFN-γ is positive, and 3 are judged as that IL-2 is positive, IL-10 wholes
For feminine gender.There is no the IFN-γ and IL-2 factors checks of a sample while being judged to the positive.
3) in 10 Healthy People samples, IFN-γ and the IL-2 factors are all shown as feminine gender, have 2 to be judged as that IL-10 is positive
Property.
According to result above, tested and analyzed with reference to IFN-γ and IL-2 the two cell factors and can judge work
Dynamic property tuberculosis sample, sensitivity is high, specificity is good, if adding other factor auxiliary judgments can not effectively improve detection
Rate, while can also increase the risk of erroneous judgement.
At present, due to not being specifically designed for the detection method of active tuberculosis, clinician is substantially by multiple inspections
Looking into index carries out comprehensive descision to diagnose, therefore the detection method accuracy in the present invention is high, and clinician can be helped quick
Judge active tuberculosis (can also judge with reference to other a small amount of index comprehensives), and in the inventive method, be judged to the sample of feminine gender
Further can also carry out the detection of additive method and finally determine result.
Above-described embodiment is the present invention preferably embodiment, but embodiments of the present invention not by above-described embodiment
Limit, other any Spirit Essences without departing from the present invention and the change, modification, replacement made under principle, combine, simplification,
Equivalent substitute mode is should be, is included within protection scope of the present invention.
Claims (6)
1. the reagent of quantitative determination tuberculosis correlation factor IFN-γ and IL-2 detects the application in tuberculosis in vitro.
2. application according to claim 1, it is characterised in that：Quantitative detecting reagent includes enzyme linked immunosorbent assay, enzyme-linked
Reagent used by immune spot-ing, chemoluminescence method, liquid chip method.
3. application according to claim 1, it is characterised in that：Tuberculosis is active tuberculosis.
4. a kind of method of vitro detection active tuberculosis, it is characterised in that：Using differential stimulus antigen in vitro inducing peripheral
Blood mononuclear cell so as to produce tuberculosis correlation factor, by the secretory volume of detection wherein IFN-γ and IL-2, judgement sample is
It is no for active tuberculosis, the method is not used in medical diagnosis on disease.
5. method according to claim 4, it is characterised in that：Specific antigen includes ESAT-6, CFP-10 and Rv1985c
At least one of three kinds of albumen, form are selected from fusion protein, protein mixture or the peptide fragment of these three albumen.
6. method according to claim 4, it is characterised in that：Detection method used includes enzyme linked immunosorbent assay, enzyme
Connection immune spot-ing, chemoluminescence method, liquid chip method.
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| CN201610925713.8A CN106546737A (en) | 2016-10-24 | 2016-10-24 | A kind of method of vitro detection active tuberculosis |
| PCT/CN2016/105469 WO2018076404A1 (en) | 2016-10-24 | 2016-11-11 | Method for in vitro detection of active tuberculosis |
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| CN201610925713.8A CN106546737A (en) | 2016-10-24 | 2016-10-24 | A kind of method of vitro detection active tuberculosis |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| 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 |
| RU2687553C1 (en) * | 2018-03-26 | 2019-05-15 | Федеральное государственное бюджетное учреждение "Омский аграрный научный центр" (ФГБНУ "Омский АНЦ") | Method of lifetime differential diagnosis of bovine tuberculosis and mycobacterioses |
| CN110702918A (en) * | 2019-09-24 | 2020-01-17 | 广州迪澳医疗科技有限公司 | Kit for rapidly detecting active tuberculosis |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| RU2702609C1 (en) * | 2018-08-06 | 2019-10-08 | Федеральное государственное бюджетное учреждение "Новосибирский научно-исследовательский институт туберкулеза" Министерства здравоохранения Российской Федерации (ФГБУ "ННИИТ" Минздрава России) | Method for simulating tuberculosis infection in vitro |
| 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 |
| CN115418347A (en) * | 2022-08-31 | 2022-12-02 | 广东医科大学 | TG model and construction method |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103604933A (en) * | 2013-11-27 | 2014-02-26 | 华中科技大学同济医学院附属同济医院 | Kit for detecting active tuberculosis based on antigen specificity TNF-alpha-ELISA and application thereof |
| CN104004069A (en) * | 2014-06-06 | 2014-08-27 | 上海交通大学医学院 | Reagent for mycobacterium tuberculosis infection detection, clinical treatment effect tracking and antituberculous vaccine development and application thereof |
| CN104459129A (en) * | 2015-01-05 | 2015-03-25 | 复旦大学附属华山医院 | Diagnostic kit for distinguishing active and latent mycobacterium tuberculosis infection |
| CN105131125A (en) * | 2015-09-11 | 2015-12-09 | 广州迪澳医疗科技有限公司 | Fusion protein for inducing peripheral blood mononuclear cells (PBMC) to generate tuberculosis-related cytokines |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| SI2315773T1 (en) * | 2008-07-25 | 2016-12-30 | Glaxosmithkline Biologicals S.A. | Polypeptides, polynucleotides and compositions for use in the treatment of latent tubeculosis |
| SG182573A1 (en) * | 2010-01-27 | 2012-08-30 | Glaxosmithkline Biolog Sa | Modified tuberculosis antigens |
| KR20150058152A (en) * | 2012-07-10 | 2015-05-28 | 트랜스진 에스아이 | Mycobacterial antigen vaccine |
| CN105601747B (en) * | 2015-10-21 | 2019-05-28 | 中山大学 | A kind of tubercle bacillus fusion protein and its application in induction peripheral blood mononuclear cells generation cell factor |
| CN105541975A (en) * | 2015-12-29 | 2016-05-04 | 广州迪澳医疗科技有限公司 | Mixed polypeptide for producing tuberculosis associated cell factors by inducing peripheral blood mononuclear cell |
-
2016
- 2016-10-24 CN CN201610925713.8A patent/CN106546737A/en active Pending
- 2016-11-11 WO PCT/CN2016/105469 patent/WO2018076404A1/en active Application Filing
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103604933A (en) * | 2013-11-27 | 2014-02-26 | 华中科技大学同济医学院附属同济医院 | Kit for detecting active tuberculosis based on antigen specificity TNF-alpha-ELISA and application thereof |
| CN104004069A (en) * | 2014-06-06 | 2014-08-27 | 上海交通大学医学院 | Reagent for mycobacterium tuberculosis infection detection, clinical treatment effect tracking and antituberculous vaccine development and application thereof |
| CN104459129A (en) * | 2015-01-05 | 2015-03-25 | 复旦大学附属华山医院 | Diagnostic kit for distinguishing active and latent mycobacterium tuberculosis infection |
| CN105131125A (en) * | 2015-09-11 | 2015-12-09 | 广州迪澳医疗科技有限公司 | Fusion protein for inducing peripheral blood mononuclear cells (PBMC) to generate tuberculosis-related cytokines |
Non-Patent Citations (5)
| Title |
|---|
| ELENA CHIAPPINI ET AL: "Potential Role of M. tuberculosis Specific IFN-γ and IL-2 ELISPOT Assays in Discriminating Children with Active or Latent Tuberculosis", 《PLOS ONE》 * |
| MAHO SUZUKAWA ET AL: "Combined Analysis of IFN-γ, IL-2, IL-5, IL-10, IL-1RA and MCP-1 in QFT Supernatant Is Useful for Distinguishing Active Tuberculosis from Latent Infection", 《PLOS ONE》 * |
| PAULIN N. ESSONE ET AL: "Bifunctional T-Cell-Derived Cytokines for the Diagnosis of Tuberculosis and Treatment Monitoring", 《RESPIRATION》 * |
| R. BISELLI ET AL: "Detection of interleukin-2 in addition to interferon-γ discriminates active tuberculosis patients, latently infected individuals, and controls", 《CLINICAL MICROBIOLOGY AND INFECTION》 * |
| 胡忠义: "《实验结核病学》", 28 February 2014 * |
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| 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 |
| 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 |
| RU2687553C1 (en) * | 2018-03-26 | 2019-05-15 | Федеральное государственное бюджетное учреждение "Омский аграрный научный центр" (ФГБНУ "Омский АНЦ") | Method of lifetime differential diagnosis of bovine tuberculosis and mycobacterioses |
| CN110702918A (en) * | 2019-09-24 | 2020-01-17 | 广州迪澳医疗科技有限公司 | Kit for rapidly detecting active tuberculosis |
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