CN106124767A - Respiratory syncytial virus IgA antibody test strip and detection method thereof - Google Patents
Respiratory syncytial virus IgA antibody test strip and detection method thereof Download PDFInfo
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- CN106124767A CN106124767A CN201610318961.6A CN201610318961A CN106124767A CN 106124767 A CN106124767 A CN 106124767A CN 201610318961 A CN201610318961 A CN 201610318961A CN 106124767 A CN106124767 A CN 106124767A
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- syncytial virus
- respiratory syncytial
- iga antibody
- saliva
- test strip
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- 229920001220 nitrocellulos Polymers 0.000 claims abstract description 7
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6854—Immunoglobulins
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56983—Viruses
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- 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
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Abstract
A kind of respiratory syncytial virus 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 syncytial virus f-protein antigenic polypeptide fragments.The present invention gathers saliva sample by sterile absorbent cotton, then is extruded by saliva sample syringe, by the IgA antibody in respiratory syncytial virus IgA antibody flash chromatography paper slip detection saliva, it is achieved quick diagnosis patient infects the purpose of respiratory syncytial virus.This method is simple to operate, and result is quick, is suitable for medical institutions at different levels quick diagnosis respiratory syncytial virus infection.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 syncytial virus and diagnose popularization rate clinically.
Description
Technical field
The invention belongs to field of medical examination, specifically disclose a kind of respiratory syncytial virus IgA antibody Test paper
Bar and detection method thereof.
Background technology
Respiratory syncytial virus (RSV is called for short syncytial virus, also belongs to Paramyxoviridae) is to cause infantile viral pneumonia
Common cause of disease, can cause interstitial pneumonia, and bronchiolitis.Respiratory syncytial virus can be changed point according to molecular biology
It is 2 kinds of hypotypes, A type and Type B.Respiratory syncytial virus is propagated through the air spittle and close contact, incubation period 3～7 days.Infant
Symptom is heavier, can have high heat, rhinitis, pharyngitis and laryngitis, show as bronchiolitis and pneumonia later.Minority sick child can concurrently in
Otitis, pleuritis and myocarditis etc..Primary disease is more common in infant, and the most more than half is baby within 1 years old, and man is more than female, its
Ratio is about 1.5～2:1.Initial stage visible cough, nasal obstruction, majority of cases has high heat, the highest can be to 41 DEG C, the high heat time is most
Being 1～4 day, minority is 5～8 days.Mild cases dyspnea and nervous symptoms do not write, in, serious symptom have significantly breathe tired
Difficult, breathe heavily suppress, cyanotic lips, flaring nares and three depressions sign, minority severe cases also can Complicated by Heart Failure.Syncytial virus lung nearly ten years
Scorching and bronchiolitis accounts for China's Infant Viral Pneumonia first, its symptom and parainfluenza virus pneumonia, mild influenza
Virus bronchopneumonia and mild adenovirus pneumonia the most almost cannot be distinguished from, but respiratory syncytial virus and parainfluenza virus, stream
Influenza Virus, the Therapeutic Method of adenovirus have bigger difference.The specificity medication of respiratory syncytial virus is ribavirin and handkerchief
Profit pearl monoclonal antibody.It is generally available ribavirin (ribavirin) nebulae inhalation, short-term heavy dose nebulae inhalation syncytial virus infection
Effectively.In early days respiratory syncytial virus is diagnosed and treatment is had the biggest value, hospital stays and the expense of patient can be reduced
With, reduce the medical burden of society.
The method of early diagnosis of respiratory syncytial virus is more, the most in a organized way cell culture technology, serology, molecule life
Three art of thing.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.Cell separation cultivation is the goldstandard that respiratory syncytial virus infection determines
Method, but owing to time length (more than 5 days), experiment condition require that high, operator's technology requires that the shortcomings such as height limit it
In clinical diagnostic applications.The detection of serology IgM antibody ELISA and serum IgM antibody immune fluoroscopic examination are that clinical practice is wider
Method, by detection serum in respiratory syncytial virus IgM antibody understand patient's recent infection virus situation, clinic is examined
Control certain help.But a lot of respiratory syncytial virus patients are less than the child of 3 years old, the blood vessel of more child is relatively thin,
And blood drawing is had fear, therefore bring certain difficulty to clinical staff operation.Another antibody mediated immunity fluoroscopic examination needs
Want the personnel of specialty and special fluorescence microscope, which has limited it at clinical Generalization Ability.PCR detection of nucleic acids is respiratory tract
The syncytial virus method that detection method medium sensitivity is the highest in early days, sick by the respiratory syncystial in detection patient's bottleneck throat cell
Poison judges infection conditions.This technology needs technical professional and special fluorescent PCR amplification instrument, and primary care department is more difficult
Carry out this diagnostic method.Fluorescence PCR assay needs to gather patient's bottleneck throat cell, is easier to cause patient's discomfort to cause anthostele
Or vomiting occurs.Respiratory syncytial virus is belonging to lower respiratory infection, therefore gathers brush,throat and does not sometimes collect disease
The cell that poison infects, causes detection leakage phenomenon.
When respiratory syncytial virus invades human body by respiratory system, human immune system starts to start first and prevents
Line, is to carry out epidemic prevention by mucosal immunity to stop respiratory syncytial virus in the field planting of human body.Mucosal immunity mainly passes through saliva
Liquid produces the IgA antibody opposing virus infection of virus-specific, and therefore virus IgA antibody produces and can indicate Virus Infection
Occur.The present invention is respiratory syncytial virus IgA antibody in detection patient's saliva, is a kind of noninvasive detection technique, easily quilt
Infant accepts.And using flash chromatography method, the detection time, less than 30 minutes, can facilitate primary care department and scene
Detection, improves the work efficiency of medical personnel, symptomatic treatment as early as possible, shortens Rehabilitation process.
Summary of the invention
For the problem overcoming prior art to exist, the present invention provides a kind of respiratory syncytial virus IgA antibody detection examination
Paper slip and detection method thereof.
Present invention firstly provides a kind of respiratory syncytial virus IgA antibody test strip, including sample pad, labelling
Pad, nitrocellulose filter, absorbent paper, backboard, described cellulose membrane includes that being coated with respiratory syncytial virus f-protein resists
The detection line of former polypeptide fragment and the control line of people's IgA antibody.
Further, the aminoacid sequence such as sequence table of described respiratory syncytial virus f-protein 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, the saliva sample of described ELISA test strip people.
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.
Present invention also offers and utilize described ELISA test strip respiratory syncytial virus IgA antibody method: it is to be measured right to gather
As 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.
Present disclosure also includes the acquisition method of a kind of people's saliva sample: the sterile absorbent cotton bundled by cotton thread is placed
In object Sublingual to be measured, after absorption saliva, squeeze out detection specimen with syringe again.
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 syncytial virus IgA antibody flash chromatography paper slip detection saliva, it is achieved quick diagnosis patient infects respiratory syncystial
The purpose of virus.This method is simple to operate, and result is quick, is suitable for medical institutions at different levels quick diagnosis respiratory syncytial virus sense
Dye.Collect specimen is without invasive, it is to avoid traditional blood sampling and collection Nasopharyngeal swabs bring discomfort to patient, also improve medical personnel
Work efficiency, improve respiratory syncytial virus diagnose 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 syncytial virus 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 syncytial virus IgA antibody of the present invention.
Fig. 5 is the negative schematic diagram of ELISA test strip respiratory syncytial virus IgA antibody of the present invention.
Fig. 6 is the invalid schematic diagram of ELISA test strip respiratory syncytial virus IgA antibody of the present invention.
Detailed description of the invention
Prepared by [embodiment 1] respiratory syncytial virus 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 syncytial virus f-protein antigenic polypeptide fragments is Ser Val Ile Thr Ser Leu
Gly Ala Ile Val Ser Cys Tyr Gly Lys Thr Lys Cys Thr Ala Ser Asn Lys Asn Arg
Gly Ile Ile Lys Thr Phe Ser Asn Gly Cys Asp Tyr Val Ser Asn Lys Gly Val(Seq
ID NO.1), 43 aminoacid altogether, entrust gill biochemical corp, Shanghai to be responsible for synthesis, purity is more than 98%, and 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 syncystial virus F protein antigenic polypeptide fragments, wherein people's IgA antibody
Concentration is 1.5mg/ml, and respiratory syncystial virus F protein antigenic polypeptide fragments concentration is 1mg/ml.Such as accompanying drawing 1, incite somebody to action both uniformly
Spraying on NC film, wherein people's IgA antibody is sprayed onto location of C on NC film, and respiratory syncystial virus F protein antigenic polypeptide fragments is sprayed onto
T location on NC film.The NC film sprayed is placed on humidity 10-30%, under the conditions of temperature 20-35 DEG C drying and processing 12 hours with
On, 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 syncytial virus 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 syncytial virus IgA antibody in saliva sample, can be with the goat-anti people's IgA antibody glue in label pad
Body gold label (C line) combines, respiratory syncystial virus F protein 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 syncytial virus IgA antibody, sheep in specimen
Anti-human IgA antibody colloid gold label thing can cross detection line (T line), continues to be up to control line and ties with people's IgA antibody (C line)
Close, control line is formed red stripes (accompanying drawing 5).If control line (C line) is formed without red stripes, illustrate that test strips is lost
Effect, result unreliable (accompanying drawing 6).
[embodiment 5] respiratory syncytial virus IgA of the present invention 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 syncystial respectively
Virus IgA antibody test strip diagnostic method is tested with fluorescence PCR method, result such as table 1:
The respiratory syncytial virus IgA antibody test strip diagnostic method of table 1 present invention compares knot with fluorescence PCR method
Really
Respiratory syncytial virus 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 (10)
1. a respiratory syncytial virus 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 syncytial virus f-protein antigenic polypeptide fragments
Detection line and the control line of people's IgA antibody.
Respiratory syncytial virus IgA antibody test strip the most according to claim 1, it is characterised in that described breathing
The aminoacid sequence of road syncytial virus f-protein antigenic polypeptide fragments is as described in sequence table Seq ID NO.1.
Respiratory syncytial virus 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 syncytial virus IgA antibody test strip the most according to claim 3, it is characterised in that described mark
Note 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 syncytial virus IgA antibody test strip described in any bar in claim 1-4, its feature exists
In, the saliva sample of described ELISA test strip people.
Respiratory syncytial virus IgA antibody test strip the most according to claim 5, it is characterised in that described saliva
Specimen 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. utilize the detection respiratory syncytial virus IgA antibody method 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.
10. the acquisition method of people's saliva sample, it is characterised in that it is to be measured right to be positioned over by the sterile absorbent cotton that cotton thread bundlees
As Sublingual, after absorption saliva, squeeze out detection specimen with syringe again.
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| CN110487999A (en) * | 2019-09-06 | 2019-11-22 | 上海菲恒生活用品有限公司 | A kind of easy influenza virus detector |
| CN111707834A (en) * | 2020-06-24 | 2020-09-25 | 深圳海博生物技术有限公司 | IgA antibody detection kit for various respiratory pathogens |
| CN113354734A (en) * | 2021-07-14 | 2021-09-07 | 北京保图生物技术有限公司 | Kit for rapidly detecting viruses and preparation method thereof |
| CN119192307A (en) * | 2024-10-08 | 2024-12-27 | 北京溯本源和生物科技有限公司 | A respiratory syncytial virus polypeptide antigen and its application |
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Application publication date: 20161116 |