CN108267589B - Application of mycobacterium tuberculosis protein in preparing product for diagnosing latent tuberculosis infected person and/or active tuberculosis - Google Patents
Application of mycobacterium tuberculosis protein in preparing product for diagnosing latent tuberculosis infected person and/or active tuberculosis Download PDFInfo
- Publication number
- CN108267589B CN108267589B CN201611259564.2A CN201611259564A CN108267589B CN 108267589 B CN108267589 B CN 108267589B CN 201611259564 A CN201611259564 A CN 201611259564A CN 108267589 B CN108267589 B CN 108267589B
- Authority
- CN
- China
- Prior art keywords
- protein
- tuberculosis
- ala
- gly
- leu
- 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.)
- Active
Links
Images
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
- G01N33/6854—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
The invention provides application of mycobacterium tuberculosis protein in preparing products for diagnosing latent tuberculosis infectors and/or active tuberculosis. The method provided by the invention can simultaneously detect the level of serum antibody response of three different mycobacterium tuberculosis proteins, so that latent tuberculosis infectors and active tuberculosis patients can be better identified and diagnosed, the sensitivity is higher, and the specificity is better.
Description
Technical Field
The invention relates to the field of biomedicine, in particular to application of mycobacterium tuberculosis protein in preparing a product for diagnosing latent tuberculosis infectors and/or active tuberculosis.
Background
Tuberculosis caused by mycobacterium tuberculosis is still one of diseases seriously threatening human health, according to the latest report of the world health organization, 1040 ten thousands of new tuberculosis cases are found in 2015 all the world, about 180 thousands of tuberculosis cases die from tuberculosis, 91.8 thousands of new tuberculosis cases and 3.8 thousands of tuberculosis cases are found in China, and the total number of the tuberculosis cases is the fourth position in the world. The work of preventing and treating tuberculosis is still far in the world or in China, and the infection rate of tubercle bacillus is still high especially in some underdeveloped countries with poor sanitary conditions.
After a human body is infected with mycobacterium tuberculosis, about only 10% of people develop active tuberculosis, and 90% of people do not develop the active tuberculosis, but the tubercle bacillus in the human body is not completely eliminated, but is latent in the human body, namely tuberculosis latent infection, the people have no clinical symptoms, most of people do not develop the active tuberculosis in the life, and only 5% -10% of people can cause the tuberculosis when the immunity of the human body is reduced. At present, 1/3 people all over the world infect tubercle bacillus, and the number of visible latent infected people is large, so that the latent infected people need to be monitored, and when the infection state changes, the drug therapy is timely given to achieve early discovery and early treatment, so that the disease deterioration can be prevented, the treatment time and the side effect caused by the drug therapy can be reduced, and the spreading of tuberculosis can be reduced. Therefore, the detection and identification of the two states after tubercle bacillus infection are very important for the prevention and treatment of tuberculosis.
At present, the detection methods of tubercle bacillus infection comprise a plurality of methods, such as Tuberculin Skin Test (TST), gamma-interferon release test (IGRA), traditional phlegm acid-fast bacterium smear, rapid culture of mycobacteria, amplification (quantitative and qualitative) of tuberculosis specific nucleic acid fragments and the like. TST is skin allergy, has long service time, is mainly used for outpatient screening, although the test is simple and easy to implement, the same result can be produced in BCG inoculation when the tubercle bacillus infection is detected, natural infection cannot be identified, the specificity is not strong, and in addition, latent tuberculosis infection and active tuberculosis cannot be identified. The gamma interferon release assay (IGRA) is mainly used for detecting cellular immune response, although the effect of BCG vaccination can be eliminated, latent infectors and active tuberculosis patients cannot be distinguished, and treated patients and current infectors cannot be distinguished. However, the traditional sputum acid-fast bacteria smear, the rapid culture of mycobacteria and the like are mainly used for detecting active tuberculosis patients, have no significance for judging latent infection states, and in addition, bacteriological evidence may not be found for the active tuberculosis patients with atypical clinical symptoms. Therefore, the method for accurately identifying latent infection and active tuberculosis is of great value.
Based on different host protective immune responses caused by tubercle bacillus infection, the method for searching the serum marker to distinguish latent infection and active tuberculosis has good application prospect. The serological diagnosis method of antigen-antibody reaction is of great interest because of its simplicity, rapidity and easy availability of specimens, and some specific antigens of mycobacterium tuberculosis have been used for serological diagnosis of tubercle bacillus infection, such as secreted protein antigens: antigen 85 complex antigens, i.e., Ag85A, Ag85B, and Ag85C, etc.; lipoprotein antigen: 16kDa, 27kDa, 38kDa antigen, etc.; glycolipid antigens: lipoarabinomannan antigens, tuberculosyl lipoid antigens, and the like; however, in actual use, the antigens have the problems of low positive detection rate, cross immunity with other mycobacteria and the like, and although the antigens have certain values in the general survey, the monitoring of treatment effect, the judgment of prognosis and the prompt of relapse of high risk groups, the antigens cannot be used for the definite diagnosis of latent mycobacterium tuberculosis infection at present.
Therefore, it is very important to find a new method and marker capable of accurately diagnosing latent tuberculosis infectors and active tuberculosis patients.
Disclosure of Invention
The invention aims to solve the technical problems that in the prior art, products for identifying and diagnosing latent tuberculosis infectors and/or active tuberculosis have low positive detection rate and cannot identify and diagnose latent mycobacterium tuberculosis infection, and provides an application of mycobacterium tuberculosis protein in preparing products for diagnosing latent tuberculosis infectors and/or active tuberculosis.
In order to solve the technical problems, the technical scheme provided by the invention on one hand is as follows:
use of a product for detecting the level of an anti-Rv 0284 antibody, an anti-Rv 2002 antibody and/or an anti-Rv 2421c antibody in a sample in the manufacture of a product for identifying, diagnosing and/or screening latent tuberculosis infection and/or active tuberculosis.
In the technical scheme of the invention, any product for detecting the level of the anti-Rv 0284 antibody, the anti-Rv 2002 antibody and/or the anti-Rv 2421c antibody in the sample can be used for preparing a product for identifying, diagnosing and/or screening latent tuberculosis infection and/or active tuberculosis.
Preferably, the product for detecting the level of the anti-Rv 0284 antibody, the anti-Rv 2002 antibody and/or the anti-Rv 2421c antibody in the sample comprises an Rv0284 protein, an Rv2002 protein and/or an Rv2421c protein.
More preferably, the Rv0284 protein is a protein shown in SEQ ID NO.1, or one or more amino acid residues in the amino acid sequence shown in SEQ ID NO.1 are modified to have the same function as the protein shown in SEQ ID NO. 1; the Rv2002 protein is shown in SEQ ID NO.2, or the protein which has the same function with the protein shown in SEQ ID NO.2 after one or more amino acid residues in the amino acid sequence shown in SEQ ID NO.2 are modified; the Rv2421c protein is shown in SEQ ID NO.3, or the protein which has the same function with the protein shown in SEQ ID NO.3 after one or more amino acid residues in the amino acid sequence shown in SEQ ID NO.3 are modified.
The term "modification" as used above may be any prior art modification of proteins, including, but not limited to, substitution and/or addition and/or deletion of amino acid residues to form derivatives of the original protein.
Preferably, the detection may be any suitable detection method in the prior art, but preferably, the detection method may be an enzyme-linked immunosorbent assay, an agglutination assay, a precipitation assay, an E-rosette assay, a phagocytosis assay, an immunofluorescence assay, a colloidal gold immunochromatography assay, a spotted gold immunodiafiltration assay, and/or an electrochemiluminescence assay. More preferably, in one embodiment of the present invention, the detection method is an enzyme-linked immunosorbent assay.
Preferably, the sample may be any suitable sample obtained from the subject. Preferably, however, the sample comprises serum, plasma, saliva, urine, pleural effusion and/or cerebrospinal fluid. More preferably, in one embodiment of the present invention, the sample is serum.
Another aspect of the present invention provides a marker composition for identifying, diagnosing and/or screening latent infection with tuberculosis and/or active tuberculosis, the marker composition consisting of an anti-Rv 0284 antibody, an anti-Rv 2002 antibody and/or an anti-Rv 2421c antibody;
wherein, the Rv0284 is a protein which is shown in SEQ ID NO.1 or has the same function with the protein shown in SEQ ID NO.1 after one or more amino acid residues in the amino acid sequence shown in SEQ ID NO.1 are modified; the Rv2002 is a protein shown in SEQ ID NO.2, or one or more amino acid residues in an amino acid sequence shown in SEQ ID NO.2 are modified to have the same function with the protein shown in SEQ ID NO. 2; the Rv2421c is a protein shown in SEQ ID NO.3, or one or more amino acid residues in the amino acid sequence shown in SEQ ID NO.3 are modified to have the same function with the protein shown in SEQ ID NO. 3.
In another aspect of the invention, the invention provides a use of the above marker composition in the preparation of a product for identifying, diagnosing and/or screening latent tuberculosis infection and/or active tuberculosis.
Another aspect of the invention provides a kit for identifying, diagnosing and/or screening latent tuberculosis infection and/or active tuberculosis, the kit comprising Rv0284 protein, Rv2002 protein and/or Rv2421c protein.
Preferably, the Rv0284 protein is a protein shown in SEQ ID NO.1, or one or more amino acid residues in the amino acid sequence shown in SEQ ID NO.1 are modified to have the same function as the protein shown in SEQ ID NO. 1; the Rv2002 protein is shown in SEQ ID NO.2, or the protein which has the same function with the protein shown in SEQ ID NO.2 after one or more amino acid residues in the amino acid sequence shown in SEQ ID NO.2 are modified; the Rv2421c protein is shown in SEQ ID NO.3, or the protein which has the same function with the protein shown in SEQ ID NO.3 after one or more amino acid residues in the amino acid sequence shown in SEQ ID NO.3 are modified.
The Rv0284 protein, Rv2002 protein and/or Rv2421c protein contained in the kit can be in any form, including but not limited to direct protein samples, and the form of immobilizing the protein on a carrier, such as a detection chip.
The kit comprises at least kit instructions in addition to Rv0284 protein, Rv2002 protein and/or Rv2421c protein.
The specification describes criteria for determining the results: and (3) using the three proteins for joint detection, if at least 2 antibodies in the anti-Rv 0284 antibody, the anti-Rv 2002 antibody and/or the anti-Rv 2421c antibody are positive in the detection result of the sample to be detected, judging that the sample to be detected is positive in active tuberculosis, otherwise, judging that the sample to be detected is latent tuberculosis infection.
In another aspect, the invention provides the use of the above-mentioned kit in the preparation of a product for identifying, diagnosing and/or screening latent tuberculosis infection and/or active tuberculosis.
The invention has the beneficial effects that:
the kit can simultaneously detect the level of the serum antibody response of three different mycobacterium tuberculosis proteins, thereby better identifying and diagnosing latent tuberculosis infectors and active tuberculosis patients, and having higher sensitivity and better specificity.
Drawings
Fig. 1 is a statistical graph of ELISA for detecting the expression levels of anti-Rv 0284 antibody, anti-Rv 2002 antibody and anti-Rv 2421c antibody in sera of tuberculosis latent infected patients (n-93) and active tuberculosis patients (n-92);
FIG. 2 is a ROC graph of antibodies corresponding to the joint detection of three antigens;
DESCRIPTION OF THE SEQUENCES
SEQ ID NO.1 is the amino acid sequence of the Rv0284 protein in the embodiment of the invention;
SEQ ID NO.2 is the amino acid sequence of the Rv2002 protein in the embodiment of the invention;
SEQ ID NO.3 is an amino acid sequence of the Rv2421c protein in the embodiment of the invention.
Detailed Description
The invention discloses application of a product for detecting the level of an anti-Rv 0284 antibody, an anti-Rv 2002 antibody and/or an anti-Rv 2421c antibody in a sample in preparing a product for identifying, diagnosing and/or screening latent tuberculosis infection and/or active tuberculosis. Those skilled in the art can modify the process parameters appropriately to achieve the desired results with reference to the disclosure herein. It is expressly intended that all such alterations and modifications which are obvious to those skilled in the art are deemed to be incorporated herein by reference, and that the techniques of the invention may be practiced and applied by those skilled in the art without departing from the spirit, scope and range of equivalents of the invention.
In the present invention, unless otherwise specified, scientific and technical terms used herein have the meanings that are commonly understood by those skilled in the art.
In order to make those skilled in the art better understand the technical solution of the present invention, the following detailed description of the present invention is provided with reference to specific embodiments.
Example 1: collection and processing of samples
The sample to be detected is serum, the collection method comprises the steps of collecting blood of a subject in an empty stomach, using a common biochemical test tube without anticoagulant, standing at room temperature, centrifuging at the normal temperature of 3000 r/min for 10 minutes after natural coagulation, collecting the serum, subpackaging in a freezing tube, and placing in a low-temperature refrigerator at the temperature of minus 80 ℃ for storage for use in experiments.
The samples used in the present invention are divided into two groups: a latent infection group and an active tuberculosis patient group; wherein, the latent infection group refers to the patients without clinical chief complaints, positive TST experiment and T-SPOT experiment, and negative chest X-ray; the active tuberculosis patient group refers to confirmed tuberculosis patients which are positive through smear or culture and do not start or just start systemic chemical drug treatment; both groups excluded diabetes, HIV infection and other autoimmune diseases.
The specimens of the latent infection group used by the invention are from epidemiological research projects of epidemiological research rooms of tuberculosis breast tumor research institute of Beijing city, the specimens of the active tuberculosis patient group are from inpatients of each disease area of tuberculosis department of Beijing thoracic hospital affiliated to capital medical university, the subject is approved by the research ethics committee of Beijing thoracic hospital affiliated to capital medical university, and all research objects are informed.
Example 2: detection of samples using ELISA assays
1. Establishing various buffers, diluents, reaction solutions and stop solutions required by an ELSIA experiment:
(1) coating buffer (pH9.6, 0.05mol/L carbonate buffer):
Na2CO31.59g，NaHCO3adding double distilled water to 1000ml for 2.93g, and adjusting the pH to 9.6;
(2) PBS solution ph 7.4:
NaCl 8.0g，KCl 0.2g，KH2PO40.2g，Na2HPO4·12H2O 2.9g，
adding double distilled water to 1000ml, and adjusting the pH value to 7.4;
(3) PBST wash ph 7.4:
adding Tween-200.5 ml into 1000ml of PBS solution, and adjusting the pH to 7.4;
(4) blocking solution (PBS solution with skim milk powder, ph 7.4):
5g of skimmed milk powder is added into 100ml of PBS solution;
(5) substrate buffer: 0.2M NaH2PO4(28.4g/L)25.7ml，
24.3ml of 0.1M citric acid (19.2g/L) and 50ml of distilled water were added.
(6) TMB (tetramethylbenzidine) use solution 0.5ml substrate buffer (pH5.5) 0.5ml 0.75% H2O232μl；
(7) Stop solution (2M H)2SO4)：21.32ml H2SO4，178.68ml H2O。
(8) Secondary antibody: horseradish peroxidase (HRP) labeled goat anti-human IgG;
(9) a 96-hole enzyme label plate; nunc Incorporated (Theromo, Gebenhagen, Denmark)
2. A detection step:
(1) coating: the Rv0284, Rv2002 and Rv2421c antigens were dissolved in coating buffer (5ug/ml) respectively; adding 100ul per well into a 96-well enzyme label plate, and standing overnight at 4 ℃;
(2) washing the plate: PBST cleaning solution 300 ul/hole, 3min, wash 3 times;
(3) and (3) sealing: blocking solution, 300 ul/hole, and incubating for 1h at room temperature;
(4) washing the plate: washing solution 300 ul/hole for 3min for 3 times;
(5) adding serum: incubating the serum to be tested at room temperature for 1h at 100 ul/hole after dilution at a ratio of 1: 400;
(6) washing the plate: washing solution 300 ul/hole for 3min for 5 times;
(7) adding an enzyme-labeled secondary antibody: 100ul of freshly diluted enzyme-labeled antibody (1:30000) per well, and incubating for 1h at room temperature;
(8) washing the plate: washing solution 300 ul/hole for 3min for 5 times;
(9) color development: incubation is carried out for 5-10min at room temperature in a dark place with 100 ul/hole of TMB developing solution prepared freshly;
(10) and (4) terminating: stop solution 2M H2SO450 ul/well;
(11) and (3) determination: the microplate reader was adjusted to 450nm and the OD of each well was measured after zeroing with the first blank PBS control well.
3. Results analysis and criteria for determination:
using SPSS22.0 software to draw an ROC curve, obtaining specificity and sensitivity corresponding to different critical points according to coordinate values, then calculating a Yoden index (the Yoden index is specificity + sensitivity-1), and obtaining the critical point OD values of three antigens respectively, wherein Rv0248 is 0.3475, Rv2002 is 0.3048, and Rv2421c is 0.4173;
the results are shown in figure 1, and show that the three proteins have the function of distinguishing latent tuberculosis infection and active tuberculosis.
And (3) judging standard:
and (3) using the three proteins for joint detection, if at least 2 antibodies in the anti-Rv 0284 antibody, the anti-Rv 2002 antibody and/or the anti-Rv 2421c antibody are positive in the detection result of the sample to be detected, judging that the sample to be detected is positive in active tuberculosis, otherwise, judging that the sample to be detected is latent tuberculosis infection.
Example 3: specificity and sensitivity of method for differential diagnosis of active tuberculosis patients and latent infected patients by applying ELISA
Adopting 92 parts of serum of a patient clinically diagnosed with active tuberculosis and 93 parts of serum of a latent infected patient; the test was carried out by the ELISA method in examples 1 and 2, and the test subject was judged to be positive for active tuberculosis by the judgment criteria in example 2, as shown in FIG. 2, with the result that the assay specificity was 83.87% and the sensitivity was 79.35%. In FIG. 2, the abscissa 1-specificity represents the false positive rate, and the ordinate sensitivity represents the true positive rate.
The results prove that the kit can detect the level of the serum antibody response of three different mycobacterium tuberculosis proteins, can better identify and diagnose latent tuberculosis infectors and active tuberculosis patients, and has higher sensitivity and better specificity.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
SEQUENCE LISTING
<110> Beijing thoracic Hospital affiliated to capital medical university
<120> use of protein of mycobacterium tuberculosis in preparing products for diagnosing latent tuberculosis infection and/or active tuberculosis
In (A) use
<130>None
<160>3
<170>PatentIn version 3.5
<210>1
<211>645
<212>PRT
<213> Mycobacterium tuberculosis
<400>1
Met Val Glu Val Glu Arg His Ser Tyr Asp Val Val Val Ile Gly Ala
1 5 10 15
Gly Gly Ala Gly Leu Arg Ala Val Ile Glu Ala Arg Glu Arg Gly Leu
20 25 30
Lys Val Ala Val Val Cys Lys Ser Leu Phe Gly Lys Ala His Thr Val
35 40 45
Met Ala Glu Gly Gly Cys Ala Ala Ala Met Gly Asn Ala Asn Pro Lys
50 55 60
Asp Asn Trp Lys Thr His Phe Gly Asp Thr Met Arg Gly Gly Lys Phe
65 70 75 80
Leu Asn Asn Trp Arg Met Ala Glu Leu His Ala Lys Glu Ala Pro Asp
85 90 95
Arg Val Trp Glu Leu Glu Thr Tyr Gly Ala Leu Phe Asp Arg Thr Asp
100 105 110
Asp Gly Arg Ile Ser Gln Arg Asn Phe Gly Gly His Thr Tyr Pro Arg
115 120 125
Leu Ala His Val Gly Asp Arg Thr Gly Leu Glu Leu Ile Arg Thr Leu
130 135 140
Gln Gln Lys Val Val Ser Leu Gln Gln Glu Asp His Ala Glu Leu Gly
145 150 155 160
Asp Tyr Glu Ala Arg Ile Lys Val Phe Ala Glu Cys Thr Ile Thr Glu
165 170 175
Leu Leu Lys Asp Gln Gly Ala Ile Ala Gly Ala Phe Gly Tyr Trp Arg
180 185 190
Glu Ser Gly Arg Phe Ile Val Phe Glu Ala Pro Ala Val Val Leu Ala
195 200 205
Thr Gly Gly Ile Gly Lys Ser Phe Lys Val Thr Ser Asn Ser Trp Glu
210 215 220
Tyr Thr Gly Asp Gly His Ala Leu Ala Leu Arg Ala Gly Ala Thr Leu
225 230 235 240
Asn Met Glu Phe Val Gln Phe His Pro Thr Gly Met Val Trp Pro Pro
245 250 255
Ser Val Lys Gly Ile Leu Val Thr Glu Gly Val Arg Gly Asp Gly Gly
260 265 270
Val Leu Lys Asn Ser Glu Asn Ser Arg Phe Met Phe Asp Tyr Ile Pro
275 280 285
Pro Val Phe Lys Gly Gln Tyr Ala Glu Thr Glu Glu Glu Ala Asp Gln
290 295 300
Trp Leu Lys Asp Asn Asp Ser Ala Arg Arg Thr Pro Asp Leu Leu Pro
305 310 315 320
Arg Asp Glu Val Ala Arg Ala Ile Asn Ser Glu Val Lys Ala Gly Arg
325 330 335
Gly Thr Pro His Gly Gly Val Tyr Leu Asp Ile Ala Ser Arg Leu Thr
340 345 350
Pro Ala Glu Ile Lys ArgArg Leu Pro Ser Met Tyr His Gln Phe Lys
355 360 365
Glu Leu Ala Glu Val Asp Ile Thr Thr Gln Ala Met Glu Val Gly Pro
370 375 380
Thr Cys His Tyr Val Met Gly Gly Val Glu Val Asp Ala Asp Thr Gly
385 390 395 400
Ala Ala Thr Val Pro Gly Leu Phe Ala Ala Gly Glu Cys Ala Gly Gly
405 410 415
Met His Gly Ser Asn Arg Leu Gly Gly Asn Ser Leu Ser Asp Leu Leu
420 425 430
Val Phe Gly Arg Arg Ala Gly Leu Gly Ala Ala Asp Tyr Val Arg Ala
435 440 445
Leu Ser Ser Arg Pro Ala Val Ser Ala Glu Ala Ile Asp Ala Ala Ala
450 455 460
Gln Gln Ala Leu Ser Pro Phe Glu Gly Pro Lys Asp Gly Ser Ala Pro
465 470 475 480
Glu Asn Pro Tyr Ala Leu His Met Asp Leu Gln Tyr Val Met Asn Asp
485 490 495
Leu Val Gly Ile Ile Arg Asn Ala Asp Glu Ile Ser Arg Ala Leu Thr
500 505 510
Leu Leu Ala Glu Leu Trp Ser ArgTyr His Asn Val Leu Val Glu Gly
515 520 525
His Arg Gln Tyr Asn Pro Gly Trp Asn Leu Ser Ile Asp Leu Arg Asn
530 535 540
Met Leu Leu Val Ser Glu Cys Val Ala Arg Ala Ala Leu Gln Arg Thr
545 550 555 560
Glu Ser Arg Gly Gly His Thr Arg Asp Asp His Pro Gly Met Asp Pro
565 570 575
Asn Trp Arg Arg Ile Leu Leu Val Cys Arg Ala Thr Glu Thr Met Gly
580 585 590
Thr Gly Gly Ser Gly Ser Gly Asp Ser Asn Cys His Ile Asn Val Thr
595 600 605
Gln Gln Leu Gln Thr Pro Met Arg Pro Asp Leu Leu Glu Leu Phe Glu
610 615 620
Ile Ser Glu Leu Glu Lys Tyr Tyr Thr Asp Glu Glu Leu Ala Glu His
625 630 635 640
Pro Gly Arg Arg Gly
645
<210>2
<211>260
<212>PRT
<213> Mycobacterium tuberculosis
<400>2
Met Ser Gly Arg Leu Ile Gly Lys Val Ala Leu Val Ser Gly Gly Ala
1 5 10 15
Arg Gly Met Gly Ala Ser His Val Arg Ala Met Val Ala Glu Gly Ala
20 25 30
Lys Val Val Phe Gly Asp Ile Leu Asp Glu Glu Gly Lys Ala Val Ala
35 40 45
Ala Glu Leu Ala Asp Ala Ala Arg Tyr Val His Leu Asp Val Thr Gln
50 55 60
Pro Ala Gln Trp Thr Ala Ala Val Asp Thr Ala Val Thr Ala Phe Gly
65 70 75 80
Gly Leu His Val Leu Val Asn Asn Ala Gly Ile Leu Asn Ile Gly Thr
85 90 95
Ile Glu Asp Tyr Ala Leu Thr Glu Trp Gln Arg Ile Leu Asp Val Asn
100 105 110
Leu Thr Gly Val Phe Leu Gly Ile Arg Ala Val Val Lys Pro Met Lys
115 120 125
Glu Ala Gly Arg Gly Ser Ile Ile Asn Ile Ser Ser Ile Glu Gly Leu
130 135 140
Ala Gly Thr Val Ala Cys His Gly Tyr Thr Ala Thr Lys Phe Ala Val
145 150 155 160
Arg Gly Leu Thr Lys Ser Thr Ala Leu Glu Leu Gly Pro Ser Gly Ile
165 170 175
Arg Val Asn Ser Ile His Pro Gly Leu Val Lys Thr Pro Met Thr Asp
180 185 190
Trp Val Pro Glu Asp Ile Phe Gln Thr Ala Leu Gly Arg Ala Ala Glu
195 200 205
Pro Val Glu Val Ser Asn Leu Val Val Tyr Leu Ala Ser Asp Glu Ser
210 215 220
Ser Tyr Ser Thr Gly Ala Glu Phe Val Val Asp Gly Gly Thr Val Ala
225 230 235 240
Gly Leu Ala His Asn Asp Phe Gly Ala Val Glu Val Ser Ser Gln Pro
245 250 255
Glu Trp Val Thr
260
<210>3
<211>211
<212>PRT
<213> Mycobacterium tuberculosis
<400>3
Met Gly Gly Thr Phe Asp Pro Ile His Tyr Gly His Leu Val Ala Ala
1 5 10 15
Ser Glu Val Ala Asp Leu Phe Asp Leu Asp Glu Val Val Phe Val Pro
20 25 30
Ser Gly Gln Pro Trp Gln Lys GlyArg Gln Val Ser Ala Ala Glu His
35 40 45
Arg Tyr Leu Met Thr Val Ile Ala Thr Ala Ser Asn Pro Arg Phe Ser
50 55 60
Val Ser Arg Val Asp Ile Asp Arg Gly Gly Pro Thr Tyr Thr Lys Asp
65 70 75 80
Thr Leu Ala Asp Leu His Ala Leu His Pro Asp Ser Glu Leu Tyr Phe
85 90 95
Thr Thr Gly Ala Asp Ala Leu Ala Ser Ile Met Ser Trp Gln Gly Trp
100 105 110
Glu Glu Leu Phe Glu Leu Ala Arg Phe Val Gly Val Ser Arg Pro Gly
115 120 125
Tyr Glu Leu Arg Asn Glu His Ile Thr Ser Leu Leu Gly Gln Leu Ala
130 135 140
Lys Asp Ala Leu Thr Leu Val Glu Ile Pro Ala Leu Ala Ile Ser Ser
145 150 155 160
Thr Asp Cys Arg Gln Arg Ala Glu Gln Ser Arg Pro Leu Trp Tyr Leu
165 170 175
Met Pro Asp Gly Val Val Gln Tyr Val Ser Lys Cys Arg Leu Tyr Cys
180 185 190
Gly Ala Cys Asp Ala Gly Ala Arg Ser Thr Thr Ser Leu Ala Ala Gly
195 200 205
Asn Gly Leu
210
Claims (9)
1. Use of a product for detecting the levels of an anti-Rv 0284 antibody, an anti-Rv 2002 antibody and an anti-Rv 2421c antibody in a sample in the preparation of a product for identifying, diagnosing and/or screening latent tuberculosis infection and active tuberculosis;
the sample is serum.
2. The use of claim 1, wherein the products for detecting the levels of anti-Rv 0284, anti-Rv 2002 and anti-Rv 2421c antibodies in a sample are Rv0284, Rv2002 and Rv2421c protein.
3. The use according to claim 2, wherein the Rv0284 protein is a protein of SEQ ID No. 1; the Rv2002 protein is a protein shown as SEQ ID NO. 2; the Rv2421c protein is a protein shown in SEQ ID NO. 3.
4. Use according to any one of claims 1 to 3, wherein the detection method comprises an enzyme-linked immunosorbent assay, an agglutination assay, a precipitation assay, an E rosette assay, a phagocytosis assay, an immunofluorescence assay, a colloidal gold immunochromatography assay, a dot immunogold diafiltration assay and/or an electrochemiluminescence assay.
5. A marker composition for use in identifying, diagnosing and/or screening latent infection with tuberculosis and active tuberculosis, wherein the marker composition consists of an anti-Rv 0284 antibody, an anti-Rv 2002 antibody and an anti-Rv 2421c antibody;
wherein, Rv0284 is protein shown in SEQ ID NO. 1; the Rv2002 is a protein shown as SEQ ID NO. 2; the Rv2421c is a protein shown in SEQ ID NO. 3.
6. Use of a marker composition according to claim 5 in the manufacture of a product for the identification, diagnosis and/or screening of latent tuberculosis infection and active tuberculosis.
7. A kit for identifying, diagnosing and/or screening latent tuberculosis infection and active tuberculosis, which is characterized in that the kit comprises Rv0284 protein, Rv2002 protein and Rv2421c protein.
8. The kit according to claim 7, wherein the Rv0284 protein is a protein of SEQ ID No. 1; the Rv2002 protein is a protein shown as SEQ ID NO. 2; the Rv2421c protein is a protein shown in SEQ ID NO. 3.
9. Use of a kit according to claim 7 or 8 for the preparation of a product for the identification, diagnosis and/or screening of latent tuberculosis infection and active tuberculosis.
Priority Applications (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611259564.2A CN108267589B (en) | 2016-12-30 | 2016-12-30 | Application of mycobacterium tuberculosis protein in preparing product for diagnosing latent tuberculosis infected person and/or active tuberculosis |
| CN202010371686.0A CN111521819B (en) | 2016-12-30 | 2016-12-30 | Use of mycobacterium tuberculosis proteins in the preparation of a product for diagnosing tuberculosis latency infected persons and/or active tuberculosis |
| CN202010371675.2A CN111551741B (en) | 2016-12-30 | 2016-12-30 | Use of mycobacterium tuberculosis proteins in the preparation of a product for diagnosing tuberculosis latency infected persons and/or active tuberculosis |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611259564.2A CN108267589B (en) | 2016-12-30 | 2016-12-30 | Application of mycobacterium tuberculosis protein in preparing product for diagnosing latent tuberculosis infected person and/or active tuberculosis |
Related Child Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010371686.0A Division CN111521819B (en) | 2016-12-30 | 2016-12-30 | Use of mycobacterium tuberculosis proteins in the preparation of a product for diagnosing tuberculosis latency infected persons and/or active tuberculosis |
| CN202010371675.2A Division CN111551741B (en) | 2016-12-30 | 2016-12-30 | Use of mycobacterium tuberculosis proteins in the preparation of a product for diagnosing tuberculosis latency infected persons and/or active tuberculosis |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108267589A CN108267589A (en) | 2018-07-10 |
| CN108267589B true CN108267589B (en) | 2020-10-23 |
Family
ID=62754940
Family Applications (3)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201611259564.2A Active CN108267589B (en) | 2016-12-30 | 2016-12-30 | Application of mycobacterium tuberculosis protein in preparing product for diagnosing latent tuberculosis infected person and/or active tuberculosis |
| CN202010371686.0A Active CN111521819B (en) | 2016-12-30 | 2016-12-30 | Use of mycobacterium tuberculosis proteins in the preparation of a product for diagnosing tuberculosis latency infected persons and/or active tuberculosis |
| CN202010371675.2A Active CN111551741B (en) | 2016-12-30 | 2016-12-30 | Use of mycobacterium tuberculosis proteins in the preparation of a product for diagnosing tuberculosis latency infected persons and/or active tuberculosis |
Family Applications After (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010371686.0A Active CN111521819B (en) | 2016-12-30 | 2016-12-30 | Use of mycobacterium tuberculosis proteins in the preparation of a product for diagnosing tuberculosis latency infected persons and/or active tuberculosis |
| CN202010371675.2A Active CN111551741B (en) | 2016-12-30 | 2016-12-30 | Use of mycobacterium tuberculosis proteins in the preparation of a product for diagnosing tuberculosis latency infected persons and/or active tuberculosis |
Country Status (1)
| Country | Link |
|---|---|
| CN (3) | CN108267589B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111157745A (en) * | 2020-01-17 | 2020-05-15 | 上海交通大学 | Application of human SNRPA protein in lung cancer recurrence or metastasis monitoring |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007131291A1 (en) * | 2006-05-16 | 2007-11-22 | Proteome Systems Limited | Methods of diagnosis and treatment of m. tuberculosis infection and reagents therefore |
| CN101203239A (en) * | 2005-03-31 | 2008-06-18 | 莱顿大学医药中心 | Methods and means for diagnostics, prevention and treatment of mycobacterium infections and tuberculosis disease |
| CN101294964A (en) * | 2007-11-27 | 2008-10-29 | 复旦大学附属华山医院 | Reagent and method for detecting active tuberculosis and tuberculosis dormant infection |
| CN103698530A (en) * | 2013-11-25 | 2014-04-02 | 广东体必康生物科技有限公司 | Application of mycobacterium tuberculosis protein in preparation of products used for diagnosis of active tuberculosis |
| EP2417456B1 (en) * | 2009-04-09 | 2016-07-06 | Ajit Lalvani | Diagnostic mycobacterium tuberculosis test |
Family Cites Families (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1278769A2 (en) * | 2000-04-19 | 2003-01-29 | Statens Serum Institut | Tuberculosis antigens and methods of use thereof |
| WO2002059715A2 (en) * | 2000-12-29 | 2002-08-01 | Triad Therapeutics, Inc. | Prediction of functional and structural properties of polypeptides |
| WO2006029248A2 (en) * | 2004-09-04 | 2006-03-16 | Haper Laboratories Llc | Hollow fiber technique for in vivo study of cell populations |
| WO2008134806A1 (en) * | 2007-05-04 | 2008-11-13 | The University Of Sydney | Mycobacterium antigens |
| EP2197908A2 (en) * | 2007-09-27 | 2010-06-23 | Dako Denmark A/S | Mhc multimers in tuberculosis diagnostics, vaccine and therapeutics |
| US8361482B2 (en) * | 2007-11-27 | 2013-01-29 | Aeras Global Tb Vaccine Foundation | Recombinant BCG tuberculosis vaccine designed to elicit immune responses to mycobacterium tuberculosis in all physiological stages of infection and disease |
| US7670609B2 (en) * | 2007-11-27 | 2010-03-02 | Aeras Global Tb Vaccine Foundation | Recombinant BCG tuberculosis vaccine designed to elicit immune responses to Mycobacterium tuberculosis in all physiological stages of infection and disease |
| CN101493454A (en) * | 2008-01-21 | 2009-07-29 | 范雄林 | Tuberculosis antigen specific whole blood IFN-gamma diagnosis kit, method for producing the same and method for using same |
| CN102590502B (en) * | 2012-01-11 | 2014-07-02 | 北京市结核病胸部肿瘤研究所 | Kit for auxiliary diagnosis of tuberculosis patients |
| CN103698511B (en) * | 2013-11-25 | 2016-01-27 | 广东体必康生物科技有限公司 | The purposes of Mycobacterium tuberculosis albumen in the product of preparation diagnosis latent tuberculosis infection |
| WO2016046734A2 (en) * | 2014-09-22 | 2016-03-31 | University Of The Western Cape | Compounds and compositions for treatment of tuberculosis |
| CN105588944B (en) * | 2016-02-25 | 2017-05-24 | 首都医科大学附属北京胸科医院 | Application of AGP1, SERPINA3 and CDH1 content detection system in screening active tuberculosis patients |
-
2016
- 2016-12-30 CN CN201611259564.2A patent/CN108267589B/en active Active
- 2016-12-30 CN CN202010371686.0A patent/CN111521819B/en active Active
- 2016-12-30 CN CN202010371675.2A patent/CN111551741B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101203239A (en) * | 2005-03-31 | 2008-06-18 | 莱顿大学医药中心 | Methods and means for diagnostics, prevention and treatment of mycobacterium infections and tuberculosis disease |
| WO2007131291A1 (en) * | 2006-05-16 | 2007-11-22 | Proteome Systems Limited | Methods of diagnosis and treatment of m. tuberculosis infection and reagents therefore |
| CN101294964A (en) * | 2007-11-27 | 2008-10-29 | 复旦大学附属华山医院 | Reagent and method for detecting active tuberculosis and tuberculosis dormant infection |
| EP2417456B1 (en) * | 2009-04-09 | 2016-07-06 | Ajit Lalvani | Diagnostic mycobacterium tuberculosis test |
| CN103698530A (en) * | 2013-11-25 | 2014-04-02 | 广东体必康生物科技有限公司 | Application of mycobacterium tuberculosis protein in preparation of products used for diagnosis of active tuberculosis |
Non-Patent Citations (1)
| Title |
|---|
| Deciphering the metabolic response of Mycobacterium tuberculosis to nitrogen stress;KerstinJ.Williams，et al;《Molecular Microbiology》;20150717;第97卷(第6期);全文 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111521819B (en) | 2023-06-20 |
| CN111551741B (en) | 2023-06-16 |
| CN108267589A (en) | 2018-07-10 |
| CN111521819A (en) | 2020-08-11 |
| CN111551741A (en) | 2020-08-18 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Haque et al. | Diagnosis of amebiasis in Bangladesh | |
| JP2011095275A (en) | Detection of tuberculosis and infection by mycobacterium tuberculosis using hbha | |
| van Hooij et al. | Prototype multi-biomarker test for point-of-care leprosy diagnostics | |
| WO2000026248A2 (en) | Tuberculosis diagnostic test | |
| JP2019015737A (en) | Immunological detection method and kit for Mycoplasma pneumoniae | |
| CN108267589B (en) | Application of mycobacterium tuberculosis protein in preparing product for diagnosing latent tuberculosis infected person and/or active tuberculosis | |
| CN106939035A (en) | A kind of mycobacterium tuberculosis T cell antigen epitope polypeptide and its application | |
| McConkey S et al. | Evaluation of a rapid-format antibody test and the tuberculin skin test for diagnosis of tuberculosis in two contrasting endemic settings | |
| Abeijon et al. | Novel antigen detection assay to monitor therapeutic efficacy of visceral leishmaniasis | |
| CN108267588B (en) | Application of mycobacterium tuberculosis protein in preparing product for diagnosing latent tuberculosis infected person and/or active tuberculosis | |
| KR20170002347A (en) | Method and kit for diagnosis of tuberculosis based on Mycobacterium tuberculosis antigen-specific antibody reaction | |
| JP2019508684A (en) | Methods and kits for the diagnosis of active tuberculosis | |
| KR102132964B1 (en) | Method for Diagnosing Scrub Typhus Using Exosome derived from Orientia tsutsugamushi | |
| KR20160137007A (en) | Method and kit for TB diagnosis based on immune response to K-strain specific antigens | |
| Mosleh et al. | Serodiagnosis of cutaneous leishmaniasis in Jordan using indirect fluorescent antibody test and the enzyme-linked immunosorbent assay | |
| Chapin-Robertson et al. | Cryptococcal antigen detection from the urine of AIDS patients | |
| Sarkari et al. | A comparative study of antigen and antibody detection in visceral leishmaniasis using serum and urine-based ELISA | |
| Maleewong | Recent advances in diagnosis of paragonimiasis | |
| CN109061166B (en) | ELISpot detection kit for detecting bovine tuberculosis | |
| CN106814194B (en) | A kind of kit for distinguishing activity and latent tuberculosis infects | |
| KR20160139888A (en) | Method and kit for diagnosis of tuberculosis based on Mycobacterium tuberculosis antigen-specific antibody reaction | |
| CN116400077A (en) | Antigen composition for distinguishing active tuberculosis and latent tuberculosis and application thereof | |
| Shende et al. | A low molecular weight ES-20 protein released in vivo and in vitro with diagnostic potential in lymph node tuberculosis | |
| RU2503006C1 (en) | Diagnostic technique for tuberculosis infection | |
| CN109030825A (en) | Albumen Rv2031c for the infection of specific detection mycobacterium tuberculosis |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |