CN106168623A - Respiratory Tract Adenovirus IgA antibody test strip and detection method thereof - Google Patents
Respiratory Tract Adenovirus IgA antibody test strip and detection method thereof Download PDFInfo
- Publication number
- CN106168623A CN106168623A CN201610322236.6A CN201610322236A CN106168623A CN 106168623 A CN106168623 A CN 106168623A CN 201610322236 A CN201610322236 A CN 201610322236A CN 106168623 A CN106168623 A CN 106168623A
- Authority
- CN
- China
- Prior art keywords
- respiratory tract
- iga antibody
- adenovirus
- tract adenovirus
- iga
- 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.)
- Granted
Links
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
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56983—Viruses
Abstract
A kind of Respiratory Tract Adenovirus IgA antibody test strip, including sample pad, label pad, nitrocellulose filter, absorbent paper, backboard, it is characterized in that, described cellulose membrane includes detection line and the control line of people's IgA antibody being coated with Respiratory Tract Adenovirus hexon antigen polypeptide.The present invention gathers saliva sample by sterile absorbent cotton, then is extruded by saliva sample syringe, by the IgA antibody in Respiratory Tract Adenovirus IgA antibody flash chromatography paper slip detection saliva, it is achieved quick diagnosis patient infects the purpose of Respiratory Tract Adenovirus.This method is simple to operate, and result is quick, is suitable for medical institutions at different levels quick diagnosis Respiratory Tract Adenovirus and infects.Collect specimen is without invasive, it is to avoid traditional blood sampling and collection Nasopharyngeal swabs bring discomfort to patient, also improve the work efficiency of medical personnel, improve Respiratory Tract Adenovirus and diagnose popularization rate clinically.
Description
Technical field
The invention belongs to field of medical examination, specifically disclose a kind of Respiratory Tract Adenovirus IgA antibody test strip
And detection method.
Background technology
Adenovirus (adenovirus) particle diameter 60～90nm, does not has cyst membrane, and 20 body cubic symmetry, capsid is by 252
Individual gram of microgranule composition, wherein 240 shell microgranules are six adjacent bodies (Hexon), have group-specific α antigen.Adenovirus has 6 kinds
52 kinds of different serotypes, wherein there are about 1/3 and can cause such as acute respiratory infection, epidemic keratitis, acute hemorrhagic conjunctiva
The diseases such as inflammation, pharyngo conjunctivitis, acute pharyngitis, cystitis, infant fatal pneumonia.In China, within less than 5 years old, children acute is breathed
There are about 5% in tract disease to be caused by adenovirus, the pneumonia that wherein adenovirus causes is easily caused serious disease, adenovirus lung
Inflammation is possible not only to cause heart failure, respiratory failure, it is also possible to causes and gets involved outside lung, such as encephalitis, hepatic injury, myocarditis or cardiac muscle
Infringements etc., even result in death.Adenovirus Epidemic Scope is wide, and spread speed is fast, in recent years in the outburst of multiple countries and regions and
Popular, bring threat greatly to the public health security of people.In China, 3 types and 7 type adenoviruss are to cause serious breathing
The main two kind adenovirus that road infects.There is no effective vaccine at present to prevent, can only be suited the medicine to the illness by clinical early diagnosis
Treatment reduces the harm of Respiratory Tract Adenovirus.
The method of early diagnosis of Respiratory Tract Adenovirus is more, the most in a organized way cell culture technology, serology, molecular biosciences
Learn three art.What traditional method was the more commonly used includes that cell separation is cultivated, serum IgM antibody ELISA detects, serum
IgM antibody Immunofluorescence test, PCR detection of nucleic acids.It is that Respiratory Tract Adenovirus infects the goldstandard side determined that cell separation is cultivated
Method, but due to time length (more than 5 days), experiment condition require high, operator's technology require the shortcoming such as high limit its
Clinical diagnostic applications, this technology is used primarily in scientific research field at present.The detection of serology IgM antibody ELISA is that clinical practice is wider
Method, by detection serum in Respiratory Tract Adenovirus IgM antibody understand patient's recent infection virus situation, to clinical diagnosis and treatment
There is certain help.But a lot of Respiratory Tract Adenovirus patients are less than the child of 5 years old, owing to the blood vessel of more child is relatively thin,
And blood drawing is had fear, therefore bring certain difficulty to clinical staff operation, bring psychology to patient and the head of a family
Pressure and pain sensation are subject to.The application of serum IgM antibody immune fluoroscopic examination is also to be resisted by Respiratory Tract Adenovirus IgM in detection serum
Body understands the situation of patient's recent infection virus, and this technology needs the personnel of specialty and special fluorescence microscope, which has limited
It is at clinical Generalization Ability.PCR detection of nucleic acids is the Respiratory Tract Adenovirus method that detection method medium sensitivity is the highest in early days, logical
The Respiratory Tract Adenovirus crossed in detection patient nasopharynx throat cell judges infection conditions.This technology need technical professional and
Special fluorescent PCR amplification instrument, primary care department is more difficult carries out this diagnostic method.Additionally this technology needs to gather patient's throat
Portion's cell, is easier to cause patient's discomfort to cause anthostele or vomiting to occur.
When Respiratory Tract Adenovirus invades human body by respiratory system, and human immune system starts to start the first line of defence,
It is to carry out epidemic prevention by mucosal immunity to stop Respiratory Tract Adenovirus in human body cell field planting.Mucosal immunity is mainly produced by saliva
Sick poison specific IgA antibody opposing virus infection, therefore virus IgA antibody produces and can indicate that Virus Infection is sent out
Raw.The present invention is Respiratory Tract Adenovirus IgA antibody in detection patient's saliva, is a kind of noninvasive detection technique, easily by infant
Accept.And using flash chromatography method, the detection time, less than 30 minutes, can facilitate primary care department and on-the-spot inspection
Survey, improve the work efficiency of medical personnel, symptomatic treatment as early as possible, shorten Rehabilitation process.
Summary of the invention
For the problem overcoming prior art to exist, the present invention provides a kind of Respiratory Tract Adenovirus IgA antibody Test paper
Bar and detection method thereof.
Present invention firstly provides a kind of Respiratory Tract Adenovirus IgA antibody test strip, including sample pad, label pad,
Nitrocellulose filter, absorbent paper, backboard, it is many that described cellulose membrane includes being coated with Respiratory Tract Adenovirus hexon antigen
The detection line of fragments of peptides and the control line of people's IgA antibody.
Further, the aminoacid sequence such as sequence table of described Respiratory Tract Adenovirus hexon antigenic polypeptide fragments
Described in Seq ID NO.1.
Further, described sample pad, label pad, nitrocellulose filter, absorbent paper all stick on backboard.
Further, described label pad contains gold colloidal on anti-human IgA antibody labelling, and described anti-human IgA antibody includes
Goat-anti people's IgA polyclonal antibody, mouse-anti people's IgA monoclonal antibody, rabbit anti-human IgA monoclonal antibody, rabbit anti-human IgA Anti-TNF-α
Body.
Further, described ELISA test strip people's saliva sample.
Further, described saliva sample by being positioned over object Sublingual to be measured by sterile absorbent cotton, after absorption saliva again
The method squeezed out with syringe obtains.
The method that present invention also offers the detection Respiratory Tract Adenovirus IgA antibody of described test strips: gather object to be measured
Saliva sample, and utilize this saliva sample to contact with described test strips to detect.
Further, the acquisition method of described saliva sample is: sterile absorbent cotton is positioned over object Sublingual to be measured, absorption
Saliva sample is squeezed out with syringe again after saliva.
Further, described sterile absorbent cotton cotton thread bundlees.
The invention have the benefit that
The present invention gathers saliva sample by sterile absorbent cotton, then is extruded by saliva sample syringe, passes through respiratory tract
IgA antibody in adenovirus IgA antibody flash chromatography paper slip detection saliva, it is achieved quick diagnosis patient infects Respiratory Tract Adenovirus
Purpose.This method is simple to operate, and result is quick, is suitable for medical institutions at different levels quick diagnosis Respiratory Tract Adenovirus and infects.Gather
Specimen is without invasive, it is to avoid traditional blood sampling and collection Nasopharyngeal swabs bring discomfort to patient, also improve the work of medical personnel
Efficiency, improves Respiratory Tract Adenovirus and diagnoses popularization rate clinically.
In order to be more fully understood that and implement, describe the present invention below in conjunction with the accompanying drawings in detail.
Accompanying drawing explanation
Fig. 1 is the test strips structural representation that the present invention detects Respiratory Tract Adenovirus IgA antibody in saliva.
Fig. 2 is that syringe extrudes saliva schematic diagram.
Fig. 3 is that gold colloidal paper slip tests schematic diagram.
Fig. 4 is the positive schematic diagram of ELISA test strip Respiratory Tract Adenovirus IgA antibody of the present invention.
Fig. 5 is the negative schematic diagram of ELISA test strip Respiratory Tract Adenovirus IgA antibody of the present invention.
Fig. 6 is the invalid schematic diagram of ELISA test strip Respiratory Tract Adenovirus IgA antibody of the present invention.
Detailed description of the invention
Prepared by [embodiment 1] Respiratory Tract Adenovirus IgA antibody diagnostic reagent srip
1, raw material sources: goat-anti people's IgA antibody is purchased from Sigma company, people's IgA antibody thinks biotechnology purchased from Guangzhou promise
Company limited.The sequence of Respiratory Tract Adenovirus hexon antigenic polypeptide fragments is: Met Leu Leu Gly Asn Gly
Arg Tyr Val Pro Phe His Ile Gln Val Pro Gln Lys Phe Phe Ala Val Lys Asn Leu
Leu Leu Leu Pro Gly Ser Tyr Thr Tyr Glu Trp Asn Phe Arg Lys Asp Val Asn Met
Val Leu (Seq ID NO.1), 46 aminoacid altogether, entrust gill biochemical corp, Shanghai to be responsible for synthesis, purity is more than
98%, lyophilizing preserves.
2, prepared by goat-anti people IgA antibody colloid gold label pad
In pH value 6.8-7.0 solution, goat-anti people's IgA antibody forms goat-anti people's IgA antibody glue with colloid gold particle labelling
Body gold label (labelled amount is 25ug/ml), confining liquid composition is PEG20000 and BSA albumen.Colloid gold label thing is sprayed
On glass fibre membrane, dilution parameters is 30cm2/ ml, and be dried more than 12 hours under the conditions of humidity 15%, temperature 30 DEG C,
The colloid gold label pad of preparation seals standby.
3, nitrocellulose filter is coated
With being coated liquid dilution people's IgA antibody, Respiratory Tract Adenovirus hexon antigenic polypeptide fragments, wherein people IgA resists
The concentration of body is 1.5mg/ml, and Respiratory Tract Adenovirus hexon antigenic polypeptide fragments concentration is 1mg/ml.Such as accompanying drawing 1, incite somebody to action
Both uniformly spray on NC film, and wherein people's IgA antibody is sprayed onto location of C on NC film, Respiratory Tract Adenovirus hexon antigen
Polypeptide fragment is sprayed onto T location on NC film.The NC film sprayed is placed on humidity 10-30%, dries under the conditions of temperature 20-35 DEG C
Processing more than 12 hours, the NC film phonograph seal dried is standby.
4, assembling, cutting
In conjunction with accompanying drawing 1 schematic diagram, by absorbent paper 4, nitrocellulose filter 3, colloid gold label pad 2, glass fibre membrane 1 by suitable
Sequence is pasted on pvc backboard 5, then adjusts cutting cutter parameter and carries out cutting, is cut into the test strips of 4mm width.
Prepared by [embodiment 2] sterile absorbent cotton
Purchase the medical absorbent cotton ball of a diameter of 2cm, the cotton thread of a length of 8-10cm in binding.The cotton balls handled well is put
Put in pressure cooker, pressure condition 121 DEG C 30 minutes, place in baking box after high pressure and be dried, temperature 100 DEG C, 4 hours time, it is dried
After cotton balls aluminium foil bag vacuum seal.
[embodiment 3] saliva sample collection
The present invention is that sterile absorbent cotton is placed into the Sublingual of child, and cotton thread is exposed to outside mouth, sucks 3-5 minute.As attached
Fig. 2, by cotton balls 22, is placed in syringe 21, then loads onto syringe catch bar, light and slow promotion, it is thus achieved that saliva sample 23, uses
2ml tubule preserves.
[embodiment 4] sample test
Illustrate in conjunction with accompanying drawing 3, Respiratory Tract Adenovirus IgA paper slip is placed into equipped with in the tubule of saliva sample, sample pad
End in contact saliva sample, places 15-30 minute, then visual results.
If containing Respiratory Tract Adenovirus IgA antibody in saliva sample, can be with the goat-anti people's IgA antibody colloid in label pad
Gold label (C line) combines, Respiratory Tract Adenovirus hexon antigenic polypeptide fragments detection line (T line) being up on NC film
Specific bond, forms red stripes (accompanying drawing 4).If not containing corresponding Respiratory Tract Adenovirus IgA antibody, goat-anti in specimen
People's IgA antibody colloid gold label thing can cross detection line (T line), continues to be up to control line and combines with people's IgA antibody (C line),
Control line is formed red stripes (accompanying drawing 5).If control line (C line) is formed without red stripes, illustrate that test strips lost efficacy,
Result unreliable (accompanying drawing 6).
[embodiment 5] Respiratory Tract Adenovirus of the present invention IgA colloidal gold diagnosis method and fluorescence PCR method Comparative result
Choose clinical samples 60 example, gather saliva sample and the oropharyngeal swab specimen of patient respectively, use respiratory tract adenopathy respectively
Poison IgA antibody test strip diagnostic method is tested with fluorescence PCR method, result such as table 1:
The Respiratory Tract Adenovirus IgA antibody test strip diagnostic method of table 1 present invention and fluorescence PCR method comparative result
Respiratory Tract Adenovirus IgA colloidal gold diagnosis method of the present invention compares with fluorescence PCR method, and sensitivity reaches
95.00%, specificity 95.00%, the condition of Clinical detection can be met.
The invention is not limited in above-mentioned embodiment, if various changes or deformation to the present invention are without departing from the present invention
Spirit and scope, if these are changed and within the scope of deformation belongs to claim and the equivalent technologies of the present invention, then this
Bright being also intended to comprises these changes and deformation.
Claims (9)
1. a Respiratory Tract Adenovirus IgA antibody test strip, including sample pad, label pad, nitrocellulose filter, water suction
Paper, backboard, it is characterised in that described cellulose membrane includes being coated with Respiratory Tract Adenovirus hexon antigenic polypeptide fragments
Detection line and the control line of people's IgA antibody.
Respiratory Tract Adenovirus IgA antibody test strip the most according to claim 1, it is characterised in that described respiratory tract
The aminoacid sequence of adenovirus fusion protein antigenic polypeptide fragments is as described in sequence table Seq ID NO.1.
Respiratory Tract Adenovirus IgA antibody test strip the most according to claim 2, it is characterised in that described sample pad,
Label pad, nitrocellulose filter, absorbent paper all stick on backboard.
Respiratory Tract Adenovirus IgA antibody test strip the most according to claim 3, it is characterised in that described labelling
Pad is containing gold colloidal on anti-human IgA antibody labelling, and described anti-human IgA antibody includes goat-anti people's IgA polyclonal antibody, mouse-anti people
IgA monoclonal antibody, rabbit anti-human IgA monoclonal antibody, rabbit anti-human IgA polyclonal antibody.
5. according to the Respiratory Tract Adenovirus IgA antibody test strip described in any bar in claim 1-4, it is characterised in that
Described ELISA test strip people's saliva sample.
Respiratory Tract Adenovirus IgA antibody test strip the most according to claim 5, it is characterised in that described saliva is marked
This by being positioned over object Sublingual to be measured by sterile absorbent cotton, and the method squeezed out with syringe again after absorption saliva obtains.
7. the method utilizing the detection Respiratory Tract Adenovirus IgA antibody of test strips described in claim 1 or 2, it is characterised in that:
Gather object saliva sample to be measured, and utilize this saliva sample to contact with described test strips to detect..
Method the most according to claim 7, it is characterised in that the acquisition method of described saliva sample is: by sterile absorbent
Cotton is positioned over object Sublingual to be measured, squeezes out saliva sample with syringe again after absorption saliva.
Method the most according to claim 8, it is characterised in that described sterile absorbent cotton cotton thread bundlees.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610322236.6A CN106168623B (en) | 2016-05-12 | 2016-05-12 | Respiratory Tract Adenovirus IgA antibody test strip and its detection method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610322236.6A CN106168623B (en) | 2016-05-12 | 2016-05-12 | Respiratory Tract Adenovirus IgA antibody test strip and its detection method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106168623A true CN106168623A (en) | 2016-11-30 |
| CN106168623B CN106168623B (en) | 2018-04-24 |
Family
ID=57358983
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610322236.6A Active CN106168623B (en) | 2016-05-12 | 2016-05-12 | Respiratory Tract Adenovirus IgA antibody test strip and its detection method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106168623B (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111044728A (en) * | 2019-09-27 | 2020-04-21 | 广州医科大学附属第一医院 | IgM antibody colloidal gold test strip for rapidly detecting adenovirus and preparation method thereof |
| CN111999494A (en) * | 2020-08-18 | 2020-11-27 | 北京贝尔生物工程股份有限公司 | Kit for detecting adenovirus IgM antibody by magnetic particle chemiluminescence method and preparation method thereof |
| WO2021214467A1 (en) * | 2020-04-23 | 2021-10-28 | Cellbio | A diagnostic tool improvement comprising a pathogen binding molecule |
| WO2023155291A1 (en) * | 2022-02-18 | 2023-08-24 | 嘉兴康源科泰科技发展有限公司 | Test technique based on combined loading of saliva and nasal samples, and use thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080118911A1 (en) * | 2004-08-27 | 2008-05-22 | Gerhard Hermann | Test Device for the In-Vitro Diagnosis of Multi-Analyte Tests and Use Thereof |
| CN102928589A (en) * | 2012-11-15 | 2013-02-13 | 南京凯基生物科技发展有限公司 | Colloidal gold method detection test strip and reagent kit for IgG antibody of respiratory disease and preparation method of reagent kit |
| CN103267844A (en) * | 2013-05-06 | 2013-08-28 | 南京凯基生物科技发展有限公司 | Detection test paper strip of colloidal gold method for adenovirus IgM and IgG antibodies, kit and preparation method thereof |
-
2016
- 2016-05-12 CN CN201610322236.6A patent/CN106168623B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080118911A1 (en) * | 2004-08-27 | 2008-05-22 | Gerhard Hermann | Test Device for the In-Vitro Diagnosis of Multi-Analyte Tests and Use Thereof |
| CN102928589A (en) * | 2012-11-15 | 2013-02-13 | 南京凯基生物科技发展有限公司 | Colloidal gold method detection test strip and reagent kit for IgG antibody of respiratory disease and preparation method of reagent kit |
| CN103267844A (en) * | 2013-05-06 | 2013-08-28 | 南京凯基生物科技发展有限公司 | Detection test paper strip of colloidal gold method for adenovirus IgM and IgG antibodies, kit and preparation method thereof |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111044728A (en) * | 2019-09-27 | 2020-04-21 | 广州医科大学附属第一医院 | IgM antibody colloidal gold test strip for rapidly detecting adenovirus and preparation method thereof |
| WO2021214467A1 (en) * | 2020-04-23 | 2021-10-28 | Cellbio | A diagnostic tool improvement comprising a pathogen binding molecule |
| CN111999494A (en) * | 2020-08-18 | 2020-11-27 | 北京贝尔生物工程股份有限公司 | Kit for detecting adenovirus IgM antibody by magnetic particle chemiluminescence method and preparation method thereof |
| WO2023155291A1 (en) * | 2022-02-18 | 2023-08-24 | 嘉兴康源科泰科技发展有限公司 | Test technique based on combined loading of saliva and nasal samples, and use thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106168623B (en) | 2018-04-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106168623B (en) | Respiratory Tract Adenovirus IgA antibody test strip and its detection method | |
| CN102645537B (en) | Latex enhanced turbidimetric immunoassay kit for diagnosing gastric diseases or gastric cancer, preparation method thereof and application | |
| CN109781980A (en) | African swine fever virus rapid detection card and its application | |
| CN104198703A (en) | Human mycoplasma pneumoniae gold-marked silver-stained immunochromatographic assay kit and preparation method and application thereof | |
| CN102445537A (en) | Combined detection test paper of influenza A virus antigen and influenza B virus antigen and preparation method thereof | |
| CN106841604A (en) | A kind of Respirovirus quick detection kit and preparation method thereof | |
| CN104459144B (en) | A kind of PRV velogen strain and vaccine strain differentiate Test paper | |
| CN102539782A (en) | Immune chromatography test strip for detecting cystic echinococcosis and alveolar echinococcosis and preparation method | |
| CN108267577A (en) | A kind of EV71 viruses IgA antibody test strip | |
| CN106018800A (en) | Detecting device for Brucella infection | |
| CN101130765B (en) | Reagent kit for detecting syncytial virus of respiratory passage | |
| CN109142725A (en) | A kind of A type is general and the preparation method of the colloidal gold combined Test paper of H7N9 subtype influenza virus | |
| CN106124767A (en) | Respiratory syncytial virus IgA antibody test strip and detection method thereof | |
| CN112946294A (en) | Novel coronavirus 2019-nCoV antibody detection test strip and preparation method and application thereof | |
| CN106645714B (en) | EV71 virus IgA antibody test strips and its application | |
| CN112334481A (en) | Antibody pairs for rapid influenza B diagnostic testing | |
| CN102539755A (en) | Test strip for detecting influenza A virus antigen in secretion and preparation method thereof | |
| CN105974111B (en) | A kind of method of rheumatoid factor (IgM type) and other antigen-specific IgM antibody joint-detections | |
| CN101666800A (en) | Fast test strip of vesicular stomatitis virus (VSV) and preparation method thereof | |
| CN105891469B (en) | A kind of Portunus trituberculatus Miers reovirus test strip and preparation method thereof | |
| CN102608330A (en) | Immunochromatographic strip for detecting alveolar echinococcosis and preparation method thereof | |
| CN202433383U (en) | Joint inspection test paper for influenza A virus antigen and influenza B virus antigen | |
| CN110045105A (en) | Coxsack A16 virus IgA antibody quantum dot immune fluorescent chromatograph test strip and kit | |
| CN1866019A (en) | Quick detection reagent for syphilis leptospira antigen and preparation method thereof | |
| Dennis | A stable concentrated purified antigen for the immunological study of hydatid disease |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |