CN102120758B - Mycobacterium tuberculosis antibody binding peptide and application thereof - Google Patents
Mycobacterium tuberculosis antibody binding peptide and application thereof Download PDFInfo
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Abstract
The invention belongs to the technical field of medicaments and in particular relates to mycobacterium tuberculosis antibody binding peptide and application thereof. The general formula of an amino acid sequence is X1X2X3PPFS, wherein P represents proline; F represents phenylalanine; S represents serine; X1 to X3 represent any amino acid; preferably, X1 is L or F, X2 is P or H, and X3 is P or F; and L represents leucine and H represents histidine. The tuberculosis patient serum specific antibody-bound polypeptide can be used as a detection reagent in tuberculosis serological detection.
Description
Technical field
The invention belongs to medical technical field, be specifically related to a kind of polypeptide that can be combined with the tuberculosis patient serum specific antibody, and the application of this polypeptide in tuberculosis serological detects.
Background technology
Tuberculosis is the communicable disease of WHO priority control, is still at present the public health problem of serious harm people ' s health.According to pertinent data statistics, the whole world has 1/3 population to infect tubercule bacillus approximately, and existing tuberculosis patient is about 2,000 ten thousand, and annual neopathy 8,000,000～1,000 ten thousand people has 2,000,000 people to die from tuberculosis every year approximately, is the summation of other all Death of Infectious Diseases numbers.The tuberculosis epidemic situation of China is extremely severe, and existing tuberculosis infected students 500,000,000 accounts for 1/4 of the whole world, has every year 130000 people to die from tuberculosis, is other all kinds of transmissible diseases and parasitosis death toll summation 2 times.
The main problem that at present tuberculosis control exists is that patient's discovery rate is low, and curative ratio is low.Aspect diagnosis, existing inspection method all exists certain limitation, is difficult to reach fast and accurately diagnosis of tuberculosis.Therefore, strengthening the fundamental research of mycobacterium tuberculosis, seek quick, sensitive, easy, practical diagnosis of tuberculosis novel method, improve the recall rate of tuberculosis patient, is that present tuberculosis research needs the urgent problem that solves.
Serological testing is one of present tuberculosis laboratory three large diagnostic methods, bacteriology and Bio-molecular analysis have multiple advantage relatively, its method is simple, fast, low price, do not need special large-scale test set, even basic hospital also can extensively be carried out, pulmonary tuberculosis to sputum smear negative, the outer tuberculosis of lung and the difficult diagnosis that obtains the childhood tuberculosis of sputum specimen all have important value, become the ideal chose of early discovery and diagnosis of tuberculosis, but conventional Serological testing still exists susceptibility, the shortcoming that specificity is lower has limited clinical application, and major cause is to be difficult to obtain hypersensitivity, the antigen of high specific.
The investigator has been found that and utilizes the synthetic peptide that contains the antigen immune epi-position to replace whole antigen protein, helps to reduce immunological cross-reaction, improves specificity.Researchist from Switzerland utilizes high-throughout peptide chips technology, albumen from 61 mycobacterium tuberculosis is carried out epitope resolves, found that 12 peptides can be used for difference tuberculosis and non-tuberculosis people, for providing thinking take polypeptide as the Foundation Serology test.Yet, adopt the method for peptide chips or computer simulation, can only obtain to contain the polypeptide of linear epitope, and the B cell epitope of natural antigen is mainly take conformational epitope as main, the simple polypeptide that contains linear epitope that uses can't detect the antibody of being combined with conformational epitope, affects recall rate.Unite the multiple not synthetic peptide of colinearity or conformational epitope of tuberculosis antigen that contains, might provide more effective means for serodiagnosis lungy.
The Phage display random peptide library technology is the method that allogenic polypeptide is showed in the phage particle surface, be a kind of high flux screening system of efficient molecular evolution research, in a plurality of fields such as immunology, cytobiology and drug development, obtained using very widely.At present, the random peptide library technology is widely used in the research of antigenic epitope of various molecules (comprising albumen, polysaccharide, glycolipid etc.) as a kind of very effective technology.Utilize different lengths or the random peptide library of isomorphic map not, take special antibody as target molecule, behind too much wheel phage selection, can obtain high specificity, various epi-position structures of simulation specific antigen that avidity is high and the short peptide sequence of chemical feature, comprise linear epitope, conformational epitope.Use that peptide molecule that the display technique of bacteriophage screening obtains not only can be identified specifically as haptens and in conjunction with corresponding antibodies, thereby find the antibody among the patients serum, and these polypeptide can produce in a large number by the synthetic method of gene, for the specific detection of antibody provides fast, sensitive, easy, practical novel method.
The application that the Phage display random peptide library technology is learned context of detection at antitubercle sera also has been reported, Barenholz etc. confirm in the experiment of using Phage display random peptide library technology screening tubercule bacillus glycolipid antigen fat pectinose mannosans mimic short peptide, this phage display small peptide has preferably susceptibility and specificity in serology detects, Gevorkian etc. study discovery, antigenic determinant is the decision position that causes immunne response, can be used as the substitute of antigen in the immunodiagnosis.This chamber utilizes the Phage display random peptide library technology how anti-as target molecule take the rabbit source of mycobacterium tuberculosis two species-specific antigen CE, MPT64 in earlier stage, screening has obtained 7 peptide mimic epitopess, and Preliminary detection result shows that these epitope peptides can substitute corresponding antigens fully and be used for the serology detection.But using the Phage display random peptide library technology directly screens the present home and abroad of research of tuberculosis patient serum specific antibody binding peptide and there is not yet report Chinese and English database retrievals such as (general according to tieing up) PubMed.
Summary of the invention
According to the progress of above-mentioned technical field, the invention provides a kind of polypeptide that can be combined with the tuberculosis patient serum specific antibody, the application of this polypeptide in tuberculosis serological detects also disclosed.
A kind of polypeptide that can be combined with the tuberculosis patient serum specific antibody, its aminoacid sequence general formula is: X
1X
2X
3PPFS, wherein P represents proline(Pro), and F represents phenylalanine, and S represents Serine, X
1～X
3Represent arbitrary amino acid.
Above-mentioned polypeptide of being combined with the tuberculosis patient serum specific antibody, preferred X
1Be L or F, X
2Be P or H, X
3Be P or F, wherein L represents leucine, and H represents Histidine.
Above-mentioned tuberculosis patient serum specific antibody Binding peptide can be in tuberculosis serological detects as the application of detection reagent.
The present invention screens the small peptide of with it combination take the tuberculosis patient serum IgG of purifying as target protein from the at random linear seven peptide storehouses (New England Biolabs company) of phage display.Its aminoacid sequence of the small peptide that screens is X
1X
2X
3PPFS.
Small peptide of the present invention can be by the preparation of the methods such as the routine techniques in the technology of the present invention field such as chemosynthesis.
Description of drawings
Fig. 1: SDS-PAGE detects tuberculosis patient and the healthy human IgG that purifying obtains.Wherein: 1. low molecular weight protein (LMWP) Marker; 2. tuberculosis human IgG behind the purifying; 3: healthy human IgG behind the purifying.
Fig. 2: the ELISA detected result of single phage.
Embodiment
The present invention is further illustrated below in conjunction with embodiment.
Embodiment 1: screening and the sequential analysis of tuberculosis patient serum specific antibody binding peptide.
Experiment material.
1, phage random peptide library test kit
Linear 7 peptide storehouses (the Ph.D.-7TMPhage Display Peptide Library Kit) test kit that phage surface capsid protein III shows is available from U.S. New England Biolabs company.The titre in peptide storehouse is 2 * 10
11Pfu/ml.Recipient bacterium
E.coliThe ER2738 genotype is F ' lacIq Δ (lacZ) M15 proA+B+ zzf::Tn 10 (TetR)/fhuA2 supE thi Δ (lac-proAB) Δs (hsdMS-mcrB) 5 (rk – m k – McrBC –).The full-automatic sequencing primer of DNA (96III sequencing primer): 5 '-CCCTCATAGTTAGCGTA ACG-3 ' (SEQ ID NO.1).
2, tuberculosis patient and Healthy Human Serum
47 routine tuberculosis patient serum collections are mycobacterium tuberculosis and cultivate positive from Shanghai Pulmonary Hospital's tuberculosis section outpatient service or the tuberculosis patient of being in hospital; 37 routine Healthy Human Serum collections all have the BCG (Bacille Calmette-Guerin) vaccination history from the healthy East China the Hans of Shanghai Pulmonary Hospital's Physical Check-Ups.All persons have all got rid of complication with diabetes, autoimmune disorder and HIV and have infected.Packing after the serum specimen collection ,-70 ℃ frozen.
3, chemical reagent
Bovine serum albumin (BSA) is available from U.S. Sigma company; The anti-M13 phage monoclonal antibody of HRP mark is available from Sweden Pharmacia company; Protein A/G agarose column is pleased the gram bio tech ltd available from Shanghai; TMB, Tween-20, EDTA and PEG-8000 are available from traditional Chinese medicines group.
4, the preparation of related reagent
Item layer agarose: LB substratum+1g/L MgCl26H2O+7g/L agarose, autoclaving;
Bottom LB is dull and stereotyped: LB substratum+15g/L agar powder, and autoclaving is paved plate;
PEG/NaCl: 20% (w/v) PEG-8000, 2.5mol/L NaCl。Autoclaving, room temperature storage.
TBS damping fluid: 50mmol/L Tris-HCl (pH 7.5), 150mmol/L NaCl.Autoclaving, room temperature storage.
Sodium iodide damping fluid: 10mmol/L Tris-HCl (pH 8.0), 1mmol/L EDTA, 4mol/L NaI.The room temperature lucifuge is stored.
5, key instrument
The desk-top high-speed refrigerated centrifuge of ROTINA38 (German Hettich company); Thermo MK3 microplate reader (U.S. THERMO company); HZQ-C airbath vibrator (east, Harbin connection electronic technology development corporation, Ltd.); Protein electrophoresis groove (BIO-RAD company); Full-automatic high-pressure Sterilizers (Japanese Hirayama company); Full automatic gel imaging analysis instrument (Britain Syngene company); Ultrapure water separation system (Britain USF Elga company).
1.1 selecting and the purifying of IgG of serum specimen
Select: with 47 routine tuberculosis patients and 37 routine Healthy Human Serum samples respectively with the 16KD of this chamber preparation, CE and MPT64 epitope peptide that CFP32 recombinant protein and early stage, screening obtained detect, choose detected result all negative 12 routine tuberculosis patient serum specimens screen target molecule as this, choose simultaneously both detected results all negative 12 routine Healthy Human Serum samples as anti-sieve target molecule;
Purifying: according to working instructions, respectively get the rear post of crossing of 100 μ l mixing with 12 parts of tubercular's serum specimens that Protein A/G agarose column will be picked out, tuberculosis human IgG behind the acquisition purifying, equally, 12 parts of Healthy Human Serum samples are respectively got 100 μ l mixed post, healthy human IgG behind the acquisition purifying;
Purification: SDS-PAGE detects tuberculosis patient and the healthy human IgG that purifying obtains, see Fig. 1, calculate through spectrophotometer, concentration behind the tuberculosis human IgG purifying is 1.2g/L, purity behind the purifying is more than 90%, concentration behind the healthy human IgG purifying is 1.3g/L, and the purity behind the purifying is more than 85%.
1.2 the tuberculosis patient of purifying and healthy human IgG advance screening to seven peptide storehouses
A is coated: tuberculosis patient and healthy human IgG are coated with 96 hole enzyme plates with 100 μ g/ml respectively, 100 μ l/ holes, 4 ℃ of coated spending the night;
The b sealing: the coating buffer in the mobile hole, then with the coated 300 μ l/ holes, hole of 5 mg/ml BSA sealing, 4 ℃ of sealing 2 h;
The c washing: mobile confining liquid, use TBST(0.1%Tween-20) wash 6 times;
The healthy human IgG combination of d: dilute former generation phage library with TBST, add the coated hole 100 μ l of healthy human IgG, room temperature is shaken gently and is hatched 60 min;
The combination of e tuberculosis human IgG: draw and the unconjugated phage supernatant of healthy human IgG, add the coated hole of tuberculosis human IgG, room temperature is shaken gently and is hatched 60 min;
The coated hole of the healthy human IgG of f is cleaned: according to above-mentioned purging method, clean the coated hole of healthy human IgG 10 times with TBST;
The healthy human IgG of the non-specific wash-out of g is in conjunction with phage: with 100 μ l/ hole elution buffers (1 mg/ml BSA, 0.2 mol/L Glycine-HCl, pH 2.2) room temperature jog 10 ~ 15 min wash-outs;
H neutralization: the sucking-off elutriant, add and contain 15 μ l Tris-HCl(l mol/L, pH 9.1) in the centrifuge tube of neutralization buffer, make elutriant closely neutral;
The coated hole of i tuberculosis human IgG is cleaned: mobile unconjugated phage peptide library, according to above-mentioned purging method, wash 10 times with TBST;
The non-specific wash-out tuberculosis of j human IgG also neutralizes in conjunction with phage: with 100 μ l/ hole elution buffer room temperature jog 10 ~ 15 min wash-outs, add the neutralization buffer neutralization;
The amplification of k titer determination and phage: take out 10 μ l neutralizers and dilute by 10-1～10-4 with the LB substratum, carry out the mensuration of phage titre, repeating step a-j carries out the next round screening behind remaining amplification, the purifying.
Annotate: in order to obtain the specific binding phage of high-affinity, in next one, increase the concentration (can be increased to 0.5% from 0.1%) of Tween-20 among the washings TBST.
1.3 the amplification of phage and purifying
The single bacterium colony of picking Host Strains ER2738 is inoculated in the LB substratum 37 ℃ of high vibration overnight incubation.
The bacterium that spends the night is inoculated in the 20ml LB substratum by 1:100, adds phage to be amplified, and 4.5h is cultivated in 37 ℃ of joltings;
Bacterium liquid is transferred in the 50ml centrifuge tube 4 ℃, the centrifugal 10min of 10000r/m; Supernatant is transferred in another centrifuge tube, and recentrifuge changes 80% of supernatant in another centrifuge tube over to, adds the PEG/NaCl of 1/6 volume, mixing, and 4 ℃ of precipitations are spent the night.Then 4 ℃, the centrifugal 15min of 10000r/m abandon supernatant;
Precipitation is resuspended among the l ml TBS, then moves in the 1.5ml centrifuge tube, and 4 ℃, the centrifugal 5min of 10000r/m make the residual cells precipitation.
Supernatant moves in another 1.5ml centrifuge tube, adds the PEG/ NaCl of 1/6 volume again, precipitates 1h in 4 ℃ behind the mixing.Then 4 ℃, the centrifugal 10min of 10000r/m;
Precipitation is resuspended among the 100 μ l TBS that contain 0.02% NaN3 4 ℃, the centrifugal l min of 10000r/m.Get supernatant, 4 ℃ of storages are for subsequent use.
1.4 the mensuration of phage titre
The single bacterium colony of picking Host Strains ER2738 is inoculated in the LB cultivation, and 37 ℃ are acutely shaken to logarithmic phase (OD600 ≈ 0.5);
37 ℃ of pre-temperature LB are dull and stereotyped;
Divide after top-agar melts in microwave oven to install to sterile test tube, every pipe 3ml is incubated in 50 ℃ of water-baths;
With the LB substratum phage of wash-out is diluted by 10 times of gradient series;
The phage diluent of getting the different gradients of 10 μ l adds respectively in the ER2738 bacterium liquid of 200 μ l logarithmic phases vortex mixing, incubated at room 5min;
Behind rapid vortex in the above-mentioned mixed solution adding 3ml top-agar, bed board on the LB of 37 ℃ of preheatings flat board after the placement cooling, is inverted overnight incubation for 37 ℃ in the incubator immediately;
Calculate the plaque number: in flat board, select 100 plaque left and right sides persons to count, multiply by the number that extension rate namely gets the contained phage of per 10 μ l stostes, usually represent with pfu/ μ l or pfu/ml.
The phage of eluriating generally speaking, is with 10
1-10
4Doubly dilution, the phage after the amplification is with 10
8-10
11Doubly dilution.
1.5 the preparation of phage single-chain DNA, sequencing
The ER2738 bacterium that spends the night is inoculated in the LB substratum by 1:100, and the 1ml/ pipe is sub-packed in the culture test tube, and the picking plaque adds wherein, and 4.5-5h are cultivated in 37 ℃ of joltings;
With the phage liquid after the 500 μ l amplification, the centrifugal l 0min of 10000r/m gets the supernatant repeated centrifugation once;
Supernatant moves in another centrifuge tube, adds 200 μ lPEG/NaCl, mixing, and room temperature is placed 10min;
The centrifugal l 0min of 10000r/m abandons supernatant, and is of short duration centrifugal again, exhausts supernatant;
With the abundant resuspended precipitation of 100 μ l iodide damping fluids, add 250 μ l ethanol, incubated at room temperature 10 min.4 ℃, centrifugal 10 min of 10000r/m, abandon supernatant, with 70% washing with alcohol precipitation, with 30 μ l TE dissolution precipitations, send company to check order and be translated as aminoacid sequence, (with oppositely-96 far-end primers order-checkings of M13) adopts DNASTAR software that short peptide sequence is compared.
1.6 the ELISA of phage clone and antibody binding activity detects
Coated: tuberculosis patient and healthy human IgG are with the coated 96 hole enzyme plates of 100 μ g/ml, and other establishes the blank that only adds coating buffer, 100 μ l/ holes, 4 ℃ of coated spending the night;
Sealing: the coating buffer in the mobile hole, then with the coated hole of 5 mg/ml BSA sealing and 300 μ l/ holes, blank hole, 4 ℃ of sealing 2 h;
Washing: mobile confining liquid, use TBST(0.1%Tween-20) wash 6 times;
In conjunction with: dilute each single phage and the 1st with TBST and take turns that phage (negative control) to titre is 10 after the elutriation
8Pfu/ μ l, 100 μ/hole is added to respectively in coated hole and the blank hole, 37 ℃ of 1h;
Washing: mobile in conjunction with liquid, TBST washes plate 6 times;
Enzyme labelled antibody combination: in each reacting hole, add the anti-M13 phage antibody of the HRP mark of 1:5000 dilution, 100 μ l/ holes, 37 ℃ of l h;
Washing: mobile response liquid, TBST are washed plate 6 times;
Colour developing: add the tmb substrate colour developing, 2 mol/L H2SO4 termination reactions;
Reading: spectrophotometer is read OD
450The results are shown in Figure 2.
Embodiment 2: positive single phage is to the detection of serum specimen.
Experiment material.
Serum specimen: 47 parts of active tuberculosis patients (the sputum specimen mycobacterium tuberculosis is cultivated all positive) and 37 parts have the Healthy Human Serum sample of BCG (Bacille Calmette-Guerin) vaccination history to be Shanghai Pulmonary Hospital's serum specimen storehouse preservation.
Main agents: goat anti-human igg's antibody of HRP mark is available from U.S. Sigma company
The preparation of related reagent:
Coating buffer: 17.5 mmol/LNa2CO3,32.5 mmol/L NaHCO3,15 mmol/LMgCl26H2O, pH 9.6
PBS damping fluid: 0.02mol/L Na2HPO4,0.02mol/L NaH2PO4.
Enzyme working fluid: 3%BSA, the PBS damping fluid
Plant and instrument:
The desk-top high-speed refrigerated centrifuge of ROTINA38 (German Hettich company); Thermo MK3 microplate reader (U.S. THERMO company); HZQ-C airbath vibrator (east, Harbin connection electronic technology development corporation, Ltd.); Full-automatic high-pressure Sterilizers (Japanese Hirayama company); Full automatic gel imaging analysis instrument (Britain Syngene company); Ultrapure water separation system (Britain USF Elga company)
2.1 the preparation of positive single phage: with 1.3
2.2 phage titre is measured: with 1.4
2.3 the ELISA of serum specimen detects
Coated: positive single phage is diluted to 1 * 10 with coating buffer
8Pfu/ μ l, 100 μ l/ holes are coated with 96 hole enzyme plates, 4 ℃ of coated spending the night;
Sealing: remove the coating buffer in the hole, PBST(0.05%Tween-20) wash plate 3 times, with the coated 300 μ l/ holes, hole of 1.5% skim-milk sealing, 37 ℃ of sealing 1h;
Washing: mobile confining liquid, wash 3 times with PBST;
In conjunction with: dilute the active tuberculosis patient or BCG (Bacille Calmette-Guerin) vaccination history Healthy Human Serum is arranged to 1:400,100 μ l/ holes, 37 ℃ of 2h with PBST;
Washing: mobile in conjunction with liquid, PBST washes plate 3 times;
Enzyme labelled antibody combination: in each reacting hole, add the goat anti-human igg's antibody with the HRP mark of enzyme working fluid 1:10000 dilution, 100 μ l/ holes, 37 ℃ of l h;
Washing: mobile response liquid, PBST are washed plate 6 times;
Colour developing: add the tmb substrate colour developing, 2 mol/L H2SO4 termination reactions;
Reading: spectrophotometer is read OD
450
The result judges: with the negative contrast of the mean value of Healthy Human Serum detected value, detected result and negative control ratio 〉=2.1 are judged to be the positive.More different positive single phages are to the recall rate of serum specimen.The results are shown in Table 1.
The positive single phage of table 1 detects serum ELISA interpretation of result
Embodiment 3: the antibodies peptide is used in tuberculosis serological detects.
Experiment material.
Serum specimen: 47 parts of active tuberculosis patients (the sputum specimen mycobacterium tuberculosis is cultivated all positive) and 37 parts have the Healthy Human Serum sample of BCG (Bacille Calmette-Guerin) vaccination history to be Shanghai Pulmonary Hospital's serum specimen storehouse preservation.
3.1 polypeptide is synthetic: according to the results of serological detection of single phage, select the preferably polypeptide (called after TP18) of single phage T18 displaying of detected result, peptide sequence is LPPPPPFS, and authorized company carries out external synthetic.
3.2 polypeptide dissolving: get a part of polypeptide dry powder, accurately after the weighing, be dissolved as the solution of 10mg/ml, 4 ℃ of preservations with ultrapure water.
3.3 coated concentration is judged: with peptide concentration doubling dilution to 0.5 μ g/ml～32 μ g/ml(0.5,1,2,4,8,16,32), coated 96 hole enzyme plates respectively, 4 ℃ of coated spending the night, ELISA detects is combined activity with Serum Antibody.The result gets 4 μ g/ml as the coated concentration of the best.
3.4 serology detects
Polypeptide is diluted to 4 μ g/ml with coating buffer, and 100 μ l/ holes are coated with 96 hole enzyme plates, 4 ℃ of coated spending the night, the ELISA testing process and as a result decision method with 2.3.The results are shown in Table 2.Polypeptide has than hypersensitivity and specificity detecting of serum specimen.
Table 2 TP18 polypeptide ELISA detects as a result statistical study of serum
| Group | Positive | Negative | Recall rate |
| Tuberculosis (N=47) | 33 | 14 | 70.2% |
| Normal healthy controls group (N=37) | 3 | 34 | 91.9% |
Embodiment 4:Antibodies peptide union and recombination albumen is used in tuberculosis serological detects.
Experiment material.
Serum specimen: 78 parts of active tuberculosis patients (the sputum specimen mycobacterium tuberculosis is cultivated all positive) and 98 parts have the Healthy Human Serum sample of BCG (Bacille Calmette-Guerin) vaccination history to be Shanghai Pulmonary Hospital's serum specimen storehouse preservation.
4.1 polypeptide is synthetic and dissolving: with 3.1,3.2.
4.2 recombinant protein preparation: adopt the prokaryotic expression technology to prepare the recombinant protein of Specific Antigen of Mycobacterium Tuberculosis 16KD, concentration reaches 1.2mg/ml, and purity reaches 95%.
4.3 coated concentration is judged: with peptide concentration doubling dilution to 0.5 μ g/ml～32 μ g/ml(0.5,1,2,4,8,16,32), protein concentration doubling dilution to 0.125 μ g/ml～8 μ g/ml(0.125,0.25,0.5,1,2,4,8) coated 96 hole enzyme plates respectively, 4 ℃ of coated spending the night, ELISA detects is combined activity with Serum Antibody.Polypeptide is got 4 μ g/ml as a result, and recombinant protein is got 1 μ g/ml as the coated concentration of the best.
4.4 serology detects
TP18 polypeptide and recombinant protein 16KD are diluted to 4 μ g/ml and 1 μ g/ml with coating buffer respectively, and 100 μ l/ holes are coated with 96 hole enzyme plates, 4 ℃ of coated spending the night, the ELISA testing process and as a result decision method with 2.3.The results are shown in Table 3.Associating polypeptide and recombinant protein have than hypersensitivity and specificity detecting of serum specimen, are higher than independent polypeptide detected result.
Table 3 TP18 polypeptide union and recombination protein 16 KD ELISA detects as a result statistical study of serum
| Group | Positive | Negative | Recall rate |
| Tuberculosis (N=78) | 64 | 14 | 82.1% |
| Normal healthy controls group (N=98) | 6 | 92 | 93.9% |
＜110〉Shanghai Pulmonary Hospital
＜120〉a kind of Mycobacterium tuberculosis antibody binding peptide and application thereof
<130> 001
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 20
<212> DNA
<213>
<400> 1
ccctcatagt tagcgtaacg 20
Claims (2)
1. Mycobacterium tuberculosis antibody binding peptide is characterized in that its aminoacid sequence is: LPPPPPFS, and wherein P represents proline(Pro), and F represents phenylalanine, and S represents Serine, and L represents leucine.
2. Mycobacterium tuberculosis antibody binding peptide as claimed in claim 1 is in the application of the detection reagent that detects for the preparation of tuberculosis serological.
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| CN111285920B (en) * | 2020-03-10 | 2022-08-30 | 宁夏医科大学总医院 | Amino acid sequence and nucleotide sequence of specific binding mycobacterium tuberculosis and application thereof |
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| CN101643499A (en) * | 2009-09-17 | 2010-02-10 | 上海市肺科医院 | Binding peptide of mycobacterium tuberculosis and application thereof |
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