CN106568972A - A cytokine used for evaluating tuberculosis treatment and a kit thereof - Google Patents
A cytokine used for evaluating tuberculosis treatment and a kit thereof Download PDFInfo
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- CN106568972A CN106568972A CN201610918234.3A CN201610918234A CN106568972A CN 106568972 A CN106568972 A CN 106568972A CN 201610918234 A CN201610918234 A CN 201610918234A CN 106568972 A CN106568972 A CN 106568972A
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- 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
- G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
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- 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
- G01N2333/521—Chemokines
- G01N2333/522—Alpha-chemokines, e.g. NAP-2, ENA-78, GRO-alpha/MGSA/NAP-3, GRO-beta/MIP-2alpha, GRO-gamma/MIP-2beta, IP-10, GCP-2, MIG, PBSF, PF-4 or KC
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- 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
- G01N2333/521—Chemokines
- G01N2333/523—Beta-chemokines, e.g. RANTES, I-309/TCA-3, MIP-1alpha, MIP-1beta/ACT-2/LD78/SCIF, MCP-1/MCAF, MCP-2, MCP-3, LDCF-1or LDCF-2
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- 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
- G01N2333/705—Assays involving receptors, cell surface antigens or cell surface determinants
- G01N2333/715—Assays involving receptors, cell surface antigens or cell surface determinants for cytokines; for lymphokines; for interferons
- G01N2333/7158—Assays involving receptors, cell surface antigens or cell surface determinants for cytokines; for lymphokines; for interferons for chemokines
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/12—Pulmonary diseases
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Abstract
An application of a cytokine RANTES, as a marker, in preparation of a kit used for tuberculosis diagnosis or monitoring is provided. The cytokine RANTES is combined with a cytokine Eotaxin and a cytokine IP-10 to provide a reference for evaluating curative effect prediction of a treating scheme in the early period of treatment, thus facilitating real-time adjustment of treatment schemes, improving tuberculosis treating effects, reducing side and toxic effects of ineffective medicines and facilitating enforcement and a customized medical scheme.
Description
Technical field
The present invention relates to a kind of detection technique for evaluating medical diagnosis on disease situation, more particularly to a kind of evaluation tuberculotherapy
Cytokine, and detect the test kit of these cytokines.
Background technology
Tuberculosis (Tuberculosis, TB) be by mycobacterium tuberculosis (Mycobacterium tuberculosis,
MTB a kind of chronic infectious disease for) causing, according to the statistics of WHO in 2014, the people of pulmonary tuberculosis number 930,000 newly sends out in China, in 22 knots
India being only second in the high burden country of core disease and Indonesia occupying the 3rd, tuberculosis are China's single factor test fatality rate highests
Chronic infectious disease.It is mycobacterium tuberculosis carrier that epidemiology shows that the whole world there are about 1/3 population, wherein about 10%
Carrier can develop into active tuberculosis [2].With multiple-drug resistance tuberculosis (multidrug resistance
Tuberculosiscle bacillus, MDR-TB) popular and HIV (human immunodeficiency virus) (Human immune
Deficiency vinus, HIV) infection increase, allow the infection of mycobacterium tuberculosis to be more difficult to control to, it is achieved that tuberculosis
The premise of sick primary prevention is exactly to improve diagnostic lungy.
Existing tuberculosis laboratory diagnostic method is broadly divided into three classes：Bacteriological method, molecular biology method and
Immunological method.Bacteriodiagnosises method mainly has smear staining microscope inspection and mycobacteria separation and Culture, is tuberculosis reality
Test room diagnosis common technology, but low recall rate and separation and Culture cycle due to smear for microscopic examination are longer, cause missing inspection and
Delay treatment, increases the chance of pathophoresiss；Molecular Biological Detection is based primarily upon isothermal DNA amplification, and its sensitivity increases
Easily occur false positive while high, and be difficult to obtain the suitable sample containing mycobacteria DNA thus limit its application；Exempt from
Epidemiology diagnostic method includes tuberculin test, tuberculosis antigen and antibody detection.Mycobacterium tuberculosis are typical thin
Intracellular bacterial parasite, it is cellular immunization that human immune system resists the major way of mycobacterium tuberculosis.As cell and molecule are exempted from
Epidemiology progress of research, it was found that some cytokines relevant with tuberculosis immunity and morbidity, in organism infection tuberculosis branch
During bacillus, the immunocyte of activation can secrete the immunoreation that substantial amounts of cytokine participates in body, control in tuberculosis
During treatment, its level can also produce corresponding change, so the cytokine levels for changing not only can become tuberculosis
The mark of diagnosis, it might even be possible to tuberculotherapy process is monitored, clinical application is instructed.
IFN-γ, by adjusting body immune system with corresponding receptor binding, is being tied as a kind of special-shaped glycoprotein
The antiphthisic immune state of body can be reflected in core patient's body, and, tuberculosis branch bar related to the load bacterium amount of tulase
After bacterium infection body, T cell can be recognized and produce high-caliber IFN-γ putting up a resistance, so being currently widely used for knot
The diagnosis of core disease.For example it has been directed to gamma interferon (IFN-γ) release test (interferon-gamma release
Assay, IGRAs) commercial kit T.SPOT.TB and QuantiFERON-TB Gold In-Tube (QFT-IT).But
It is that IFN-γ is used as unique detection target in existing commercial m tuberculosis infection cellular immunization detectable, and its is quick
Perception and specificity can not meet the requirement of clinic, and on the one hand its reason may be that the detection performance of IFN-γ is not high enough,
Part research shows that the accuracy of its diagnosis of tuberculosis is relatively low.On the other hand, the high disease of heterogeneity such for tuberculosis and
Speech, the detection performance of single target has certain limitation, and Chinese scholars are also being found preferably for diagnosis of tuberculosis
Biomarker, explore the probability that diagnosed of joint cytokine profiles.
Chinese invention patent application 201210559258.6 discloses a kind of monocyte chemotactic protein-2 as Tuberculous breast
The mark of water detection, mark of the MCP-2/CCL8 albumen as Differential Diagnosiss associative BCI-algebra.
Chinese invention patent application 201510317166.0 discloses a kind of diagnosis detected based on tuberculosis specificity IL-31
The test kit of m tuberculosis infection, including IL-31 capture antibody and detection antibody, tuberculosis specific antigen polypeptide, dilution
Liquid, standard substance, nitrite ion, terminate liquid, buffer, solid carrier and shrouding film etc..
Chinese invention patent application 201410182316.7 discloses a kind of immunodiagnosis kit of active tuberculosiss,
At least one in test kit energy specific recognition cytokine I-309, MIG, the IL-8, also the cytokine for being recognized can be entered
Row quantitative analyses, it is also possible to combine the specificity to antigen of mycobacterium tuberculosis 38-KDa, 32-KDa, 14-16-KDa and Ag85B
IgG antibody carries out specific recognition.
Chinese invention patent application 201410182365.0 discloses a kind of diagnosis examination of mycobacterium tuberculosis latent infection
Agent box, at least one in test kit energy specific recognition cytokine Eotaxin-2, MIG, Eotaxin, MCSF or IL-12p70
It is individual, and quantitative analyses can be carried out to the cytokine for being recognized, can be with specific recognition antigen 38-KDa, 32-KDa, 14-
The specific IgG antibodies of 16-KDa and Ag85B.
The content of the invention
It is an object of the present invention to provide a kind of RANTES cytokines are preparing diagnosis or are monitoring as label
Application in test kit lungy.
Further object is that providing a kind of RANTES cytokines and preparing diagnosis or monitoring for label
Application in the test kit of tuberculosis treatment effect.
It is yet a further object of the present invention to provide a kind of RANTES cytokines be label in tuberculosis outcome prediction
Application in test kit.
A further object of the present invention is that to provide a kind of RANTES cytokines be label in tuberculotherapy early stage
Application in curative effect predication reagent kit.
Cytokine (cytokine) is the low molecule that immunogen, mitogen or other stimulant induction various kinds of cell are produced
Amount soluble protein, repaiies with regulation inherent immunity and adaptive immunity, hemopoietic, cell growth and injury tissue
Several functions are waited again.Cytokine can be divided into interleukin, interferon, tumor necrosis factor superfamily, colony-stimulating because
Son, chemotactic factor and somatomedin etc..
The expression of inflammatory protein 10 (IP-10), eotaxin (Eotaxin) and normal T-cell and secretion
The factor (RANTES) belongs to the cytokine of chemotactic factor family.
Interleukin (IL) is main to be produced by lymphocyte, including T lymphocytes, bone-marrow-derived lymphocyte and NK cells etc..
IL-6 belongs to a kind of interleukin.
IFN-γ belongs to one kind of interferon, according to interferon produce source it is different with structure, can be divided into IFN-α,
IFN-β and IFN-γ, they are respectively by produced by leukocyte, fibroblast and activating T cell.A variety of IFN are biological
Activity is learned essentially identical, with the effect such as antiviral, antitumor and immunomodulating.
Cytokine RANTES diagnoses or monitors the application in test kit lungy as label in preparation.
Cytokine RANTES answering in the test kit for preparing diagnosis or monitoring tuberculosis treatment effect as label
With.
Application of cytokine RANTES as label in tuberculosis curative effect predication reagent kit.
Application of cytokine RANTES as label in test kit is predicted in tuberculotherapy early efficacy.
One or more of cytokine Eotaxin and cytokine IP-10 are combined as mark with cytokine RANTES
Note thing diagnoses or monitors the application in test kit lungy in preparation.
One or more of cytokine Eotaxin and cytokine IP-10 are combined as mark with cytokine RANTES
Application of the note thing in the test kit for preparing diagnosis or monitoring tuberculosis treatment effect.
A kind of test kit, with cytokine RANTES as label.
Test kit also with cytokine Eotaxin and cytokine IP-10 one or more as label.
Test kit also includes the compositionss of detection cytokine RANTES, and the combination of detection cytokine Eotaxin
The compositionss of thing and detection cytokine IP-10.
The compositionss of detection cytokine RANTES include the capture antibody of RANTES.
The beneficial effect that technical solution of the present invention is realized：
Cytokine provided by the detection present invention and combinations thereof can be to evaluate therapeutic scheme in treatment early stage to curative effect
Offer reference is provided, is beneficial to adjustment therapeutic scheme in time, improve therapeutic effect lungy, reduce the poison of ineffective agents
Side effect, beneficial to the enforcement of personalized medicine scheme.
Description of the drawings
Fig. 1 be cytokine IFN-γ, IL-6, Eotaxin each treatment time point changing trend diagram；
Fig. 2 is the changing trend diagram of cytokine IP-10 and RANTES in each treatment time point.
Specific embodiment
Technical scheme is described in detail below in conjunction with accompanying drawing.The embodiment of the present invention is only to illustrate skill of the invention
Art scheme and it is unrestricted, although being described in detail to the present invention with reference to preferred embodiment, one of ordinary skill in the art
It should be appreciated that the technical scheme invented can be modified or equivalent, without deviating from the essence of technical solution of the present invention
God and scope, it all should cover in scope of the presently claimed invention.
The foundation that each embodiment of the invention obtains data is as follows：
Pulmonary tuberculosis group：
Experiment sample selected 83 Li Yu centers in March, 2014 suffers to the pulmonary tuberculosis for receiving antituberculosis therapy between in October, 2014
Person, male 53, female 30, EDTA anticoagulant blood plasma during collecting its treatment.Require those selected in principle the age be less than 60 one full year of life, row
Except other chronic infectious diseases (such as：Hepatitis etc.) and major disease is (such as：Tumor and HIV etc.), and before no tuberculosis are controlled
Treat history.
Health examination group：
The physical examination of healthy population of any Drug therapy is not carried out in selected 29 surroundings.
Sample collection and process：
EDTA anticoagulant tubes are collected after whole blood, and 1000 × g, 15min, 4 DEG C is centrifuged.Upper plasma is collected, in order to remove completely
Platelet and precipitate, recentrifuge 10000 × g, 10min, 4 DEG C.- 70 DEG C of preservations after sample, or subpackage are determined immediately.
Cytokine is tested：
Determining test kit isMAP Kit are purchased from Merck Millipore (Merck Mi Libo) company.This
Study and lunger's treatment 0,0.5,2,4 and the blood plasma of 6 months and the blood plasma of physical examination of healthy population are detected, detect IFN-
γ, IL-6, IP-10, five kinds of cytokine levels of eotaxin, RANTES.Cytokine detection by quantitative, in strict accordance withThe explanation of MAP Kit test kits is operated.
Statistical method：
Experimental data is analyzed with the statistical analysis softwares of SPSS Statistics 17.0, and each group of data is usedTable
Show, using Kolmogorov-Smirnov Test inspection data normalities；Cytokine concentrations are in treatment different time points and group
Between difference comparison using Repeated Measurement Data variance analyses, P<0.05 is that difference is statistically significant, further two-by-two
LSD methods, the comparison in difference between the cytokine levels and healthy control group for the treatment of phase is relatively adopted to adopt Dunnett methods.
Embodiment 1
Cytokine is in each treatment period and the level statistic result of each packet in patient's Serial plasma：
The water of IFN-γ, five cytokines of IL-6, Eotaxin, IP-10 and RANTES in 83 patient's Serial plasmas
Flat (average ± standard deviation) statistics, is grouped according to different treatment times, and every kind of factor is counted respectively in different treatment time points
Between data difference (referring to table 1, Fig. 1 and Fig. 2).
Level (average ± standard deviation) of the IFN-γ of table 1 in Serial plasma
In table 1, " * " represent with first visit (0 month) when plasma levels of cytokines concentration compared with P<0.05；" # " is represented and controlled
Treat 6 months plasma levels of cytokines concentration to compare P<0.05.
Embodiment 2
Each cytokine levels treat the Variant statistical between period and physical examination of healthy population in lunger
With the Plasma Cytokine Levels in each treatment period as a group, count whether its difference between Healthy People has
Statistical significance, in as a result showing blood plasma of the lunger during whole treatment, the water of Eotaxin, IP-10 and RANTES
Averagely there is significant difference (P with physical examination of healthy population<0.05) (table 2).Pointed out by Fig. 2 B, RANTES is treating two months always
Healthy People level is had descended to the treatment level of 6 months.Table 3 has counted the level of Eotaxin, IP-10 and RANTES and has existed
The difference of Each point in time and physical examination of healthy population, in treatment, each treatment time point is consistently higher than healthy body to the level of Eotaxin
Inspection person；The level of IP-10 is significantly higher than physical examination of healthy population when treating 2 weeks and 2 months, as the carrying out for the treatment of was at 4～6 months
Although Shi Shuiping is higher than physical examination of healthy population, difference is not statistically significant；The level of RANTES is significantly higher than when treating 2 weeks
Physical examination of healthy population, treatment 2 months afterwards its level tended to physical examination of healthy population level, no significant difference.
The each cytokine levels of table 2 treat the Variant statistical between period and physical examination of healthy population in patient
Table 3 Eotaxin, IP-10 and RANTES treats Each point in time difference respectively between physical examination of healthy population in patient
Different statistics
In table 3, " * " represents the P compared with physical examination of healthy population plasma levels of cytokines concentration<0.05.
Embodiment 3
5 kinds of cytokines before the treatment with full course of therapy 6 months after concentration change trend
Before 83 lungers treatment with full course of therapy 6 months after the plasma levels of cytokines variation tendency such as institute of table 4
Show, the table summarizes each cytokine before and after treatment and rises, declines and constant patient's number of cases, and its institute in 83 patients
Accounting example.As a result show：In rising trend more of the level of Eotaxin, the level of remaining four factor to drop to master,
Wherein there are 95% patients blood plasma's RANTES levels on a declining curve.
45 kinds of cytokines of table before the treatment after concentration change trend
From result above：
1) with the carrying out for the treatment of, the level of the level of Eotaxin, IP-10 and RANTES in patients blood plasma there occurs
Change (the P of significance<0.001), the level of IP-10 overall patient through 6 months treatment after, substantially less than first visit and treatment
Initial stage；There is significant difference with first visit level in RANTES each timing node after the treatment of totality and each packet, and
Treatment 2 months with treatment 6 months after level difference have statistical significance, point out RANTES treatment 2 weeks to 2 months phases
Between variation tendency can be used for assess curative effect of disease.
2) level of RANTES has reached afterwards Healthy People level for 2 months in treatment in patients blood plasma, points out RANTES
Treatment early stage level may be more suitable for outcome prediction.
3) 5 kinds of cytokines before the treatment with full course of therapy 6 months after, Eotaxin in 83 lunger's blood plasma
In rising trend more of level, the level of remaining four factor is dropping to master, the down ratio highest of wherein RANTES
For 95%.
Claims (10)
1. cytokine RANTES diagnoses or monitors the application in test kit lungy as label in preparation.
2. application of cytokine RANTES as label in the test kit for preparing diagnosis or monitoring tuberculosis treatment effect.
3. application of cytokine RANTES as label in tuberculosis curative effect predication reagent kit.
4. application of cytokine RANTES as label in test kit is predicted in tuberculotherapy early efficacy.
5. application according to claim 3, described cytokine RANTES also with cytokine Eotaxin and cell because
One or more of sub- IP-10 are combined.
6. a kind of test kit, it is characterised in that with cytokine RANTES as label.
7. test kit according to claim 6, it is characterised in that also with cytokine Eotaxin and cytokine IP-10
One or more be label.
8. test kit according to claim 6, it is characterised in that also including the compositionss of detection cytokine RANTES.
9. test kit according to claim 8, it is characterised in that the compositionss bag of described detection cytokine RANTES
Include the capture antibody of RANTES.
10. test kit according to claim 8, it is characterised in that also including the compositionss of detection cytokine Eotaxin
With the compositionss of detection cytokine IP-10.
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| CN201610918234.3A CN106568972A (en) | 2016-10-21 | 2016-10-21 | A cytokine used for evaluating tuberculosis treatment and a kit thereof |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113406334A (en) * | 2021-06-02 | 2021-09-17 | 浙江省人民医院 | DLBCL (digital Living chromosome binding protein) related biomarker composition, application thereof and DLBCL prognosis effect prediction model |
| CN113607955A (en) * | 2020-09-16 | 2021-11-05 | 广州中医药大学顺德医院(佛山市顺德区中医院) | Novel coronavirus pneumonia staging and curative effect evaluation cytokine marker and application thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2013175459A2 (en) * | 2012-05-25 | 2013-11-28 | Stellenbosch University | Method for diagnosing tuberculosis disease |
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2016
- 2016-10-21 CN CN201610918234.3A patent/CN106568972A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2013175459A2 (en) * | 2012-05-25 | 2013-11-28 | Stellenbosch University | Method for diagnosing tuberculosis disease |
Non-Patent Citations (2)
| Title |
|---|
| J.F. DJOBA SIAWAYA ET AL: "Differential cytokine secretion and early treatment response in patients with pulmonary tuberculosis", 《CLINICAL AND EXPERIMENTAL IMMUNOLOGY》 * |
| MENG RUI LEE ET AL: "Plasma Biomarkers Can Predict Treatment Response in Tuberculosis Patients: A Prospective Observational Study", 《MEDICINE》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113607955A (en) * | 2020-09-16 | 2021-11-05 | 广州中医药大学顺德医院(佛山市顺德区中医院) | Novel coronavirus pneumonia staging and curative effect evaluation cytokine marker and application thereof |
| CN113406334A (en) * | 2021-06-02 | 2021-09-17 | 浙江省人民医院 | DLBCL (digital Living chromosome binding protein) related biomarker composition, application thereof and DLBCL prognosis effect prediction model |
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Application publication date: 20170419 |