CN108267588A - Purposes of the mycobacterium tuberculosis protein in diagnosis of tuberculosis latent infection person and/or active tuberculosis product is prepared - Google Patents

Purposes of the mycobacterium tuberculosis protein in diagnosis of tuberculosis latent infection person and/or active tuberculosis product is prepared Download PDF

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Publication number
CN108267588A
CN108267588A CN201611257494.7A CN201611257494A CN108267588A CN 108267588 A CN108267588 A CN 108267588A CN 201611257494 A CN201611257494 A CN 201611257494A CN 108267588 A CN108267588 A CN 108267588A
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China
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seq
albumen
amino acid
tuberculosis
ala
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CN201611257494.7A
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CN108267588B (en
Inventor
曹树辉
陈艳清
李传友
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Beijing Chest Hospital
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Beijing Chest Hospital
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Priority to CN202010371676.7A priority Critical patent/CN111551742B/en
Priority to CN202010371840.4A priority patent/CN111521820B/en
Priority to CN201611257494.7A priority patent/CN108267588B/en
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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
    • G01N33/6854—Immunoglobulins
    • 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
    • G01N2469/00—Immunoassays for the detection of microorganisms
    • G01N2469/20—Detection of antibodies in sample from host which are directed against antigens from microorganisms
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

Purposes of the mycobacterium tuberculosis protein in diagnosis of tuberculosis latent infection person and/or active tuberculosis product is prepared.The present invention provides detect purposes of the product of sample moderate resistance Rv0389 antibody, anti-Rv1408 antibody and/or anti-Rv2097c antibody levels in preparation discriminating, diagnosis and/or screening tuberculosis latent infection and/or active tuberculosis product.Method provided by the invention can detect the level of three different mycobacterium tuberculosis protein serum antibody responses simultaneously, and so as to preferably antidiastole tuberculosis latent infection person and active tuberculosis patient, sensitivity higher, specificity is more preferable.

Description

Mycobacterium tuberculosis protein is preparing diagnosis of tuberculosis latent infection person and/or activity Purposes in pulmonary tuberculosis product
Technical field
The present invention relates to biomedicine field, more particularly to mycobacterium tuberculosis protein is preparing diagnosis of tuberculosis latent infection Purposes in person and/or active tuberculosis product.
Background technology
The tuberculosis as caused by mycobacterium tuberculosis is still one of disease of serious threat human health, is defended according to the world The newest report of raw tissue, the new hair cases of tuberculosis in the whole world in 2015 is 10,400,000 people, and about 1,800,000 people die of tuberculosis, Chinese Neopathy number is 91.8 ten thousand people, and death toll is 3.8 ten thousand people, and morbidity total number of persons occupies the 4th, the world.Either in the whole world Or in China, preventing and controlling lungy are still to shoulder heavy responsibilities, particularly in some poor undeveloped states of sanitary condition Family, the infection rate of tulase are still very high.
After human infection mycobacterium tuberculosis, only about 10% people develops into active tuberculosis, and 90% people It does not fall ill, but internal tulase is not removed thoroughly, but hides in human body, referred to as tuberculosis latent infection, These people do not have any clinical symptoms, and most people is not fallen ill in life at it, and the only people of 5%--10% is in immunity of organisms During decline, generation lungy can be caused.There are about 1/3 people in the whole world at present to have infected tulase, it is seen that latent infection person people Number is numerous, it is therefore necessary to and to latent infection, person is monitored, and drug therapy is timely given when Infection Status changes, Accomplish early discovery, early treatment, this can not only prevent the deterioration of the state of an illness, reduce secondary work caused by the time for the treatment of and medication treatment With, moreover it is possible to reduce lungy send out.Therefore, the two states after tubercle bacillus affection detection and differentiate to prevention lungy There is very important effect with treatment.
There are many planting, tuberculin skin test (TST), gamma interferon discharge the detection method of tubercle bacillus affection at present Test (IGRA), traditional phlegm acid fast bacteria smear, mycobacteria fast culture, the amplification of tuberculosis specific nucleic acid segment (it is quantitative with It is qualitative) etc..TST is skin allergic reaction, and usage time is long, is mainly used for outpatient service screening, although experiment is simple and easy to do, inspection When surveying the situation of tubercle bacillus affection, BCG vaccination will appear identical as a result, can not differentiate natural infection, and specificity is not strong, In addition it can not also differentiate tuberculosis latent infection and active tuberculosis.Gamma interferon release test (IGRA) mainly detects cell Immune response, although the influence of BCG vaccination can be excluded, can not equally distinguish latent infection person and active tuberculosis patient, It is even more impossible to differentiate patient and Current Infection person after treating.And traditional phlegm acid fast bacteria smear, mycobacteria fast culture etc. are main To be used for detecting active tuberculosis patient, it is nonsensical for the judgement of latent infection state, further for clinical symptoms not allusion quotation The active tuberculosis patient of type possibly can not find bacteriology evidence.Therefore find one kind can accurately differentiate latent infection and The method of active tuberculosis has substantial worth.
It is different based on host-protective immune reaction caused after tubercle bacillus affection, it is latent for distinguishing to find blood serum designated object Volt infection and active tuberculosis have good application prospect.The serological diagnostic method of antigen-antibody reaction, due to its simplicity Property, agility, sample is easily obtained, and is concerned, now have some mycobacterium tuberculosis specific antigen be used for The serodiagnosis of tubercle bacillus affection, such as secreted protein antigen：85 complex antigens of antigen, i.e., Ag85A, Ag85B and Ag85C, etc.；Lipoprotein antigen：16kDa, 27kDa, 38kDa antigen etc.；Sugared lipid antigen：Fat arabian mannan resists Original, tuberculosaccharide lipoid antigen etc.；But these antigens are low there are positive rate in actual use, with other branch bars Bacterium is there are the problems such as cross-immunity, although having in people at highest risk's generaI investigation, therapeutic effect monitoring, judging prognosis and prompting recurrence There is certain value, but still cannot be used for making a definite diagnosis for latent tuberculosis mycobacterial infections at present.
Therefore, a kind of new method for being capable of Accurate Diagnosis tuberculosis latent infection person and active tuberculosis patient is found And marker is particularly important.
Invention content
The technical problem to be solved by the present invention is to be directed to antidiastole tuberculosis latent infection person and/or activity in the prior art The phthisical product of property there are positive rate is low, can not antidiastole the problem of going out latent tuberculosis mycobacterial infections, carry For a kind of mycobacterium tuberculosis protein in diagnosis of tuberculosis latent infection person and/or active tuberculosis product is prepared On the way.
In order to solve the above-mentioned technical problem, the technical solution of one aspect of the present invention offer is：
It is prepared by the product of detection sample moderate resistance Rv0389 antibody, anti-Rv1408 antibody and/or anti-Rv2097c antibody levels Differentiate, the purposes in diagnosis and/or screening tuberculosis latent infection and/or active tuberculosis product.
In the inventive solutions, arbitrary detection sample moderate resistance Rv0389 antibody, anti-Rv1408 antibody can be used And/or the product of anti-Rv2097c antibody levels is used to prepare discriminating, diagnosis and/or screening tuberculosis latent infection and/or activity Phthisical product.
Preferably, above-mentioned detection sample moderate resistance Rv0389 antibody, anti-Rv1408 antibody and/or anti-Rv2097c antibody water Rv0389 albumen, Rv1408 albumen and/or Rv2097c albumen are included in flat product.
It is highly preferred that the Rv0389 albumen is the amino acid sequence shown in SEQ ID NO.1 or shown in SEQ ID NO.1 Albumen shown in the engineered rear and SEQ ID NO.1 of one or several amino acid residues in row has the protein of identical function； The Rv1408 albumen is one or several ammonia in the amino acid sequence shown in SEQ ID NO.2 or shown in SEQ ID NO.2 Albumen has the protein of identical function after base acid residue is engineered and shown in SEQ ID NO.2；The Rv2097c albumen is After one or several amino acid residues in amino acid sequence shown in SEQ ID NO.3 or shown in SEQ ID NO.3 are engineered There is the protein of identical function with albumen shown in SEQ ID NO.3.
Term " transformation " described above can use it is arbitrary in the prior art to the remodeling method of protein, including but it is unlimited In substitution and/or addition and/or missing to amino acid residue, so as to form the derivative of original protein.
Preferably, the detection can be detection method any appropriate in the prior art, but preferably, the inspection The method of survey can be enzyme-linked immunosorbent assay, aggegation experiment, precipitation experiments, E garlands are tested, small phagocytosis is tested, immune glimmering Light experiment, colloidal gold immunochromatographimethod experiment, dot immuno gold filtration assay experiment and/or electrochemiluminescence assay.It is highly preferred that at this In one embodiment of invention, the method for the detection is enzyme-linked immunosorbent assay.
Preferably, the sample can be any appropriate sample obtained from detected object.But preferably, institute It states sample and includes serum, blood plasma, saliva, urine, Pleural effusions and/or cerebrospinal fluid.It is highly preferred that in one embodiment of the present invention In formula, the sample is serum.
Another aspect of the present invention there is provided one kind for differentiate, diagnose and/or screening tuberculosis latent infection and/or The mark compositions of active tuberculosis, the mark compositions are by anti-Rv0389 antibody, anti-Rv1408 antibody and/or resist Rv2097c antibody forms；
Wherein, the Rv0389 is one in the amino acid sequence shown in SEQ ID NO.1 or shown in SEQ ID NO.1 Albumen has the protein of identical function after a or several amino acid residues are engineered and shown in SEQ ID NO.1；It is described Rv1408 is one or several amino acid residues in the amino acid sequence shown in SEQ ID NO.2 or shown in SEQ ID NO.2 Albumen has the protein of identical function after engineered and shown in SEQ ID NO.2；The Rv2097c is SEQ ID NO.3 institutes Show or SEQ ID NO.3 shown in amino acid sequence in one or several amino acid residues it is engineered after and SEQ ID NO.3 Shown albumen has the protein of identical function.
Another aspect of the present invention differentiates, diagnoses and/or sieves in preparation there is provided a kind of above-mentioned mark compositions The purposes to come to an end in core latent infection and/or active tuberculosis product.
Another aspect of the present invention there is provided one kind for differentiate, diagnose and/or screening tuberculosis latent infection and/or The kit of active tuberculosis, the kit include Rv0389 albumen, Rv1408 albumen and/or Rv2097c albumen.
Preferably, the Rv0389 albumen is the amino acid sequence shown in SEQ ID NO.1 or shown in SEQ ID NO.1 Albumen shown in the engineered rear and SEQ ID NO.1 of one or several amino acid residues in row has the protein of identical function； The Rv1408 albumen is one or several ammonia in the amino acid sequence shown in SEQ ID NO.2 or shown in SEQ ID NO.2 Albumen has the protein of identical function after base acid residue is engineered and shown in SEQ ID NO.2；The Rv2097c albumen is After one or several amino acid residues in amino acid sequence shown in SEQ ID NO.3 or shown in SEQ ID NO.3 are engineered There is the protein of identical function with albumen shown in SEQ ID NO.3.
Rv0389 albumen, Rv1408 albumen and/or the Rv2097c albumen included in mentioned reagent box can be arbitrary shape Formula, including but not limited to direct protein example and by proteinaceous solid due to certain carrier, such as the form of detection chip.
Other than Rv0389 albumen, Rv1408 albumen and/or Rv2097c albumen, the kit at least further includes examination Agent box specification.
The criterion of result described in the specification：Using three kinds of albumen joint-detections, if the detection of sample to be tested As a result in, in anti-Rv0389 antibody, anti-Rv1408 antibody and/or anti-Rv2097c antibody, at least 2 kinds of antibodies positives are then sentenced The fixed sample to be tested is the active tuberculosis positive, is otherwise tuberculosis latent infection.
Another aspect of the present invention is there is provided a kind of mentioned reagent box in preparation discriminating, diagnosis and/or screening tuberculosis Purposes in latent infection and/or active tuberculosis product.
Beneficial effects of the present invention are：
The present invention can detect the level of three different mycobacterium tuberculosis protein serum antibody responses simultaneously, so as to Preferably antidiastole tuberculosis latent infection person and active tuberculosis patient, sensitivity higher, specificity are more preferable.
Description of the drawings
Fig. 1 is detected for ELISA in tuberculosis latent infection person (n=93) and active tuberculosis patient (n=92) serum The statistical chart of anti-Rv0389 antibody, anti-Rv1408 antibody and anti-Rv2097c antibody expressions
Fig. 2 is the ROC curve figure of the corresponding antibody of three antigens of joint-detection；
Sequence explanation
SEQ ID NO.1 are the amino acid sequence of the Rv0389 albumen in the embodiment of the present invention；
SEQ ID NO.2 are the amino acid sequence of the Rv1408 albumen in the embodiment of the present invention；
SEQ ID NO.3 are the amino acid sequence of the Rv2097c albumen in the embodiment of the present invention.
Specific embodiment
The invention discloses a kind of detection sample moderate resistance Rv0389 antibody, anti-Rv1408 antibody and/or anti-Rv2097c antibody Purposes of the horizontal product in preparation discriminating, diagnosis and/or screening tuberculosis latent infection and/or active tuberculosis product. Those skilled in the art can use for reference present disclosure, be suitably modified technological parameter realization.It is it is important to note that all similar Replacement and change it is apparent to those skilled in the art, they are considered as being included in the present invention, and phase Pass personnel can significantly be modified content described herein on the basis of the content of present invention, spirit and scope are not departed from or suitably Change is with combining, to realize and using the technology of the present invention.
In the present invention, unless otherwise stated, Science and Technology noun used herein has art technology The normally understood meaning of personnel institute.
In order to which those skilled in the art is made to more fully understand technical scheme of the present invention, with reference to specific embodiment pair The present invention is described in further detail.
Embodiment 1：The collection and processing of sample
The sample that the present invention detects is serum, and collection method is acquisition subject's blood on an empty stomach, using being not added with anti-coagulants Common biochemistry test tube, is stored at room temperature after natural coagulation, is centrifuged 10 minutes with 3000 rpms of room temperature, is collected serum and is sub-packed in Cryopreservation tube is positioned over -80 degrees Celsius of low temperature refrigerators and preserves, and uses when testing.
The sample that the present invention uses is divided into two groups：Latent infection group and active tuberculosis patient's group；Wherein, latent infection group Refer to without clinical chief complaint, TST experiments and T-SPOT experiments are the positive, C-XF negative patient；Active tuberculosis patient Group refers to positive through smear or culture, does not start or just start cycle chemistry drug therapy makes a definite diagnosis tuberculosis patient；Two groups of crowds Exclude diabetes, HIV infection and other autoimmune diseases.
The latent infection group sample that the present invention uses comes from Beijing Tuberculosis and Thoracic Tumor Research Institute's epidemiological study Room epidemiological survey project, active tuberculosis patient organize sample and come from attached BJ Chest Science Hospital's tuberculosis section of the Capital University of Medical Sciences Each lesion inpatient, this subject have obtained BJ Chest Science Hospital of the Capital University of Medical Sciences/Beijing's tuberculosis breast tumor research The approval of the Research papers committee of institute, all equal informed consents of research object.
Embodiment 2：The detection to sample is tested using ELISA
1. establish various buffer solutions, dilution, reaction solution and the terminate liquid needed for ELSIA experiments：
(1) coating buffer solution (carbonate buffer solution of pH9.6,0.05mol/L)：
Na2CO31.59g NaHCO32.93g adds distilled water to adjust pH to 9.6 to 1000ml；
(2) PBS solution of pH7.4：
NaCl 8.0g, KCl 0.2g, KH2PO40.2g, Na2HPO4·12H2O 2.9g add distilled water to be adjusted to 1000ml PH to 7.4；
(3) the PBST cleaning solutions of pH7.4：
Tween-20 0.5ml are added in 1000mlPBS solution, adjust pH to 7.4；
(4) confining liquid (PBS solution of the pH7.4 containing skimmed milk power)：
5g skimmed milk powers are added in 100mlPBS solution；
(5) substrate buffer solution：0.2M NaH2PO4(28.4g/L) 25.7ml, 0.1M citric acid (19.2g/L) 24.3ml, adds Distilled water 50ml.
(6) TMB (tetramethyl benzidine) uses liquid:TMB (10mg/5ml absolute ethyl alcohols) 0.5ml substrate buffer solutions (pH5.5) 10ml 0.75%H2O232μl；
(7) terminate liquid (2M H2SO4)：21.32ml H2SO4, 178.68ml H2O。
(8) secondary antibody：The Goat anti-Human IgG of horseradish peroxidase (HRP) label；
(9) 96 hole elisa Plates；Nunc Incorporated(Theromo,Gebenhagen,Denmark)
2. detecting step：
(1) it is coated with：Rv0389, Rv1408 and Rv2097c antigen are dissolved separately in coating buffer solution (5ug/ml)；With 100ul/ holes are added in 96 hole elisa Plates, and 4 DEG C overnight；
(2) board-washing：PBST cleaning solution 300ul/ holes, 3min are washed 3 times；
(3) it closes：Confining liquid, 300ul/ holes are incubated at room temperature 1h；
(4) board-washing：Cleaning solution 300ul/ holes, 3min are washed 3 times；
(5) increase serum：By 1:Test serum after 400 dilutions, 100ul/ holes are incubated at room temperature 1h；
(6) board-washing：Cleaning solution 300ul/ holes, 3min are washed 5 times；
(7) add ELIAS secondary antibody：The enzyme labelled antibody (1 of diluted fresh:30000) 100ul/ holes are incubated at room temperature 1h；
(8) board-washing：Cleaning solution 300ul/ holes, 3min are washed 5 times；
(9) it develops the color：The TMB developing solutions of fresh configuration, 100ul/ holes, room temperature, which is protected from light, is incubated 5-10min；
(10) it terminates：Terminate liquid 2M H2SO4, 50ul/ holes；
(11) it measures：Microplate reader is adjusted to 450nm, and each hole OD values are surveyed after returning to zero with first blank PBS control hole.
3. interpretation of result and criterion：
Using SPSS22.0 Software on Drawing ROC curves, according to coordinate value, obtain specificity corresponding to different critical point and Then susceptibility calculates youden index (youden index=specificity+susceptibility -1), obtain the critical point OD values point of three kinds of antigen It is not Rv0389 0.291, Rv1408 0.283, Rv2097c 0.308；
The results are shown in Figure 1, the results showed that, three albumen are respectively provided with the work for distinguishing tuberculosis latent infection and active tuberculosis With.
Criterion：
Using three kinds of albumen joint-detections, if in the testing result of sample to be tested, anti-Rv0389 antibody, anti-Rv1408 antibody And/or in anti-Rv2097c antibody, at least 2 kinds of antibodies positives, then it is the active tuberculosis positive to judge the sample to be tested, no It is then tuberculosis latent infection.
Embodiment 3：Using the specific and quick of the method for ELISA antidiastole active tuberculosis patients and latent infection person Perception
Use patients serum 92 part of the clinical diagnosis for active tuberculosis, 93 parts of latent infection person serum；Utilize implementation ELISA method in example 1 and 2 is detected, and judges whether person under test is active tuberculosis by the criterion of embodiment 2 The positive, the results are shown in Figure 2, and interpretation of result specificity is 86.0%, sensibility 71.7%.Abscissa 1- specificities in Fig. 2 Represent false positive rate, ordinate sensitive representations true positive rate.
Result above proves the level of the different mycobacterium tuberculosis protein serum antibody responses of present invention detection three, can be with Preferably antidiastole tuberculosis latent infection person and active tuberculosis patient, sensitivity higher, specificity are more preferable.
The above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications also should It is considered as protection scope of the present invention.
。
SEQUENCE LISTING
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<120>Mycobacterium tuberculosis protein is preparing diagnosis of tuberculosis latent infection person and/or active tuberculosis product
In purposes
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Ile Ala Ser Met
450

Claims (10)

1. the product for detecting sample moderate resistance Rv0389 antibody, anti-Rv1408 antibody and/or anti-Rv2097c antibody levels is preparing mirror Not, diagnosis and/or screening tuberculosis latent infection and/or active tuberculosis product in purposes.
2. purposes according to claim 1, described to detect sample moderate resistance Rv0389 antibody, anti-Rv1408 antibody and/or resist The product of Rv2097c antibody levels includes Rv0389 albumen, Rv1408 albumen and/or Rv2097c albumen.
3. purposes according to claim 2, which is characterized in that the Rv0389 albumen be shown in SEQ ID NO.1 or Shown in the engineered rear and SEQ ID NO.1 of one or several amino acid residues in amino acid sequence shown in SEQ ID NO.1 Albumen has the protein of identical function；The Rv1408 albumen is shown in SEQ ID NO.2 or shown in SEQ ID NO.2 Albumen shown in the engineered rear and SEQ ID NO.2 of one or several amino acid residues in amino acid sequence has identical function Protein；The Rv2097c albumen is one in the amino acid sequence shown in SEQ ID NO.3 or shown in SEQ ID NO.3 Or albumen shown in several engineered rear and SEQ ID NO.3 of amino acid residue has the protein of identical function.
4. according to the purposes described in any one in claim 1-3, which is characterized in that the method for the detection includes enzyme-linked exempt from Epidemic disease adsorption test, aggegation experiment, precipitation experiments, the experiment of E garlands, small phagocytosis experiment, immunofluorescence experiment, colloidal gold immunochromatographimethod Experiment, dot immuno gold filtration assay experiment and/or electrochemiluminescence assay.
5. according to the purposes described in any one in claim 1-3, which is characterized in that the sample includes serum, blood plasma, saliva Liquid, urine, Pleural effusions and/or cerebrospinal fluid.
6. it is a kind of for differentiating, diagnosing and/or the mark compositions of screening tuberculosis latent infection and/or active tuberculosis, It is characterized in that, the mark compositions are made of anti-Rv0389 antibody, anti-Rv1408 antibody and/or anti-Rv2097c antibody；
Wherein, the Rv0389 be one in amino acid sequence shown in SEQ ID NO.1 or shown in SEQ ID NO.1 or Albumen has the protein of identical function after several amino acid residues are engineered and shown in SEQ ID NO.1；The Rv1408 is After one or several amino acid residues in amino acid sequence shown in SEQ ID NO.2 or shown in SEQ ID NO.2 are engineered There is the protein of identical function with albumen shown in SEQ ID NO.2；The Rv2097c is shown in SEQ ID NO.3 or SEQ Albumen shown in the engineered rear and SEQ ID NO.3 of one or several amino acid residues in amino acid sequence shown in ID NO.3 Protein with identical function.
7. a kind of mark compositions as claimed in claim 6 are preparing discriminating, diagnosis and/or screening tuberculosis latent infection And/or the purposes in active tuberculosis product.
8. a kind of for differentiating, diagnosing and/or the kit of screening tuberculosis latent infection and/or active tuberculosis, feature It is, the kit includes Rv0389 albumen, Rv1408 albumen and/or Rv2097c albumen.
9. kit according to claim 8, which is characterized in that the Rv0389 albumen be shown in SEQ ID NO.1 or Shown in the engineered rear and SEQ ID NO.1 of one or several amino acid residues in amino acid sequence shown in SEQ ID NO.1 Albumen has the protein of identical function；The Rv1408 albumen is shown in SEQ ID NO.2 or shown in SEQ ID NO.2 Albumen shown in the engineered rear and SEQ ID NO.2 of one or several amino acid residues in amino acid sequence has identical function Protein；The Rv2097c albumen is one in the amino acid sequence shown in SEQ ID NO.3 or shown in SEQ ID NO.3 Or albumen shown in several engineered rear and SEQ ID NO.3 of amino acid residue has the protein of identical function.
10. a kind of kit as claimed in claim 8 or 9 prepare differentiate, diagnosis and/or screening tuberculosis latent infection and/ Or the purposes in active tuberculosis product.
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