CN106168623B - Respiratory Tract Adenovirus IgA antibody test strip and its detection method - Google Patents
Respiratory Tract Adenovirus IgA antibody test strip and its detection method Download PDFInfo
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- CN106168623B CN106168623B CN201610322236.6A CN201610322236A CN106168623B CN 106168623 B CN106168623 B CN 106168623B CN 201610322236 A CN201610322236 A CN 201610322236A CN 106168623 B CN106168623 B CN 106168623B
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Abstract
A kind of Respiratory Tract Adenovirus IgA antibody test strip, including sample pad, label pad, nitrocellulose filter, blotting paper, backboard, it is characterized in that, the cellulose membrane includes being coated with the detection line of Respiratory Tract Adenovirus hexon antigen polypeptide and the control line of people's IgA antibody.The present invention gathers saliva sample by sterile absorbent cotton, then saliva sample is extruded with syringe, and the IgA antibody in saliva is detected by Respiratory Tract Adenovirus IgA antibody flash chromatography paper slip, realizes that quick diagnosis patient infects the purpose of Respiratory Tract Adenovirus.This method is easy to operate, as a result quickly, is adapted to medical institutions' quick diagnosis Respiratory Tract Adenovirus infection at different levels.Collect specimen is avoided traditional blood sampling and collection Nasopharyngeal swabs from bringing discomfort to patient, also improves the work efficiency of medical staff without invasive, improves Respiratory Tract Adenovirus diagnosis clinically popularization rate.
Description
Technical field
The invention belongs to field of medical examination, specifically discloses a kind of Respiratory Tract Adenovirus IgA antibody test strip
And its detection method.
Background technology
Adenovirus (adenovirus) 60~90nm of particle diameter, without cyst membrane, 20 face body cubic symmetries, capsid is by 252
A gram of particulate composition, wherein 240 shell particulates are hexon (Hexon), has group-specific α antigens.Adenovirus has 6 kinds
52 kinds of different serotypes, wherein there are about 1/3 can cause such as acute respiratory infection, epidemic keratitis, acute hemorrhagic conjunctiva
The diseases such as inflammation, pharyngoconjunctivitis, acpuei pharyngitis, cystitis, infant's fatal pneumonia.Children acute breathes below China, 5 years old
There are about 5% in tract disease is as caused by adenovirus, and wherein pneumonia caused by adenovirus easily leads to serious illness, adenovirus lung
Inflammation can not only cause heart failure, respiratory failure, can also cause and be involved outside lung, such as encephalitis, hepatic lesion, myocarditis or cardiac muscle
Infringement etc., even results in death.Adenovirus Epidemic Scope is wide, and spread speed is fast, in recent years multiple countries and regions outburst and
Prevalence, the public health security to people bring great threat.In China, 3 types and 7 type adenovirus are to cause serious breathing
Two kinds of main adenovirus of road infection.It there is no effective vaccine to be prevented at present, can only be early diagnosed, suited the medicine to the illness by clinic
Treatment reduces the harm of Respiratory Tract Adenovirus.
The method of early diagnosis of Respiratory Tract Adenovirus is more, mainly cell culture technology, serology, molecular biosciences in a organized way
Learn three field technologies.Conventional method it is more commonly used be separately cultured including cell, serum IgM antibody ELISA detection, serum
IgM antibody Immunofluorescence test, PCR detection of nucleic acids.It is the definite goldstandard side of Respiratory Tract Adenovirus infection that cell, which is separately cultured,
Method, but due to the time grown (more than 5 days), experiment condition require the shortcomings of high, operating personnel's technical requirements are high limit its
Clinical diagnostic applications, at present the technology be used primarily in scientific research field.Serology IgM antibody ELISA detections are that clinical practice is wider
Method, the situation of patient's recent infection virus is understood by detecting Respiratory Tract Adenovirus IgM antibody in serum, to clinical diagnosis and treatment
There is certain help.But many Respiratory Tract Adenovirus patients are less than the children of 5 years old, since the blood vessel of more child is thinner,
And have fear to blood drawing, therefore certain difficulty is brought to clinical staff operation, bring psychology to patient and parent
Pressure and pain sensation by.Serum IgM antibody immune fluoroscopic examination application is also to be resisted by detecting Respiratory Tract Adenovirus IgM in serum
Body understands the situation of patient's recent infection virus, which needs the personnel of specialty and special fluorescence microscope, which has limited
It is in clinical Generalization Ability.PCR detection of nucleic acids is the highest method of Respiratory Tract Adenovirus early detection method medium sensitivity, is led to
The Respiratory Tract Adenovirus crossed in detection patient's nasopharynx throat cell judges infection conditions.The technology need technical professional and
Special fluorescent PCR amplification instrument, primary care department is more difficult to carry out the diagnostic method.In addition the 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, human immune system starts to start the first line of defence,
It is to carry out epidemic prevention by mucosal immunity to prevent Respiratory Tract Adenovirus from being colonized in human body cell.Mucosal immunity is mainly produced by saliva
The sick specific IgA antibody resistance virus infection of poison, therefore viral IgA antibody generation can indicate that Virus Infection is sent out
It is 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
Receive.And be less than 30 minutes using flash chromatography method, detection time, primary care department and scene can be facilitated to examine
Survey, improve the work efficiency of medical staff, as early as possible symptomatic treatment, shorten Rehabilitation process.
The content of the invention
In order to overcome problem existing in the prior art, the present invention provides a kind of Respiratory Tract Adenovirus IgA antibody Test paper
Bar and its detection method.
Present invention firstly provides a kind of Respiratory Tract Adenovirus IgA antibody test strip, including sample pad, label pad,
Nitrocellulose filter, blotting paper, backboard, the cellulose membrane are more including 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 amino acid sequence such as sequence table of the Respiratory Tract Adenovirus hexon antigenic polypeptide fragments
Described in Seq ID NO.1.
Further, the sample pad, label pad, nitrocellulose filter, blotting paper are adhered on backboard.
Further, the label pad contains the anti-human upper colloidal gold of IgA antibody mark, and the anti-human IgA antibody includes
Goat-anti people IgA polyclonal antibodies, the anti-human IgA monoclonal antibodies of mouse, rabbit-anti people IgA monoclonal antibodies, rabbit-anti people's IgA Anti-TNF-αs
Body.
Further, the ELISA test strip people saliva sample.
Further, the saliva sample is sublingual by the way that sterile absorbent cotton is positioned over object to be measured, adsorbs after saliva again
The method squeezed out with syringe obtains.
Present invention also offers the method for the detection Respiratory Tract Adenovirus IgA antibody of the test strips：Gather object to be measured
Saliva sample, and contacted and be detected with the test strips using the saliva sample.
Further, the acquisition method of the saliva sample is：It is sublingual that sterile absorbent cotton is positioned over object to be measured, absorption
After saliva saliva sample is squeezed out with syringe again.
Further, the sterile absorbent cotton is bundled with cotton thread.
Beneficial effects of the present invention are：
The present invention gathers saliva sample by sterile absorbent cotton, then saliva sample is extruded with syringe, passes through respiratory tract
IgA antibody in adenovirus IgA antibody flash chromatography paper slip detection saliva, realizes that quick diagnosis patient infects Respiratory Tract Adenovirus
Purpose.This method is easy to operate, as a result quickly, is adapted to medical institutions' quick diagnosis Respiratory Tract Adenovirus infection at different levels.Collection
Sample is avoided traditional blood sampling and collection Nasopharyngeal swabs from bringing discomfort to patient, also improves the work of medical staff without invasive
Efficiency, improves Respiratory Tract Adenovirus diagnosis clinically popularization rate.
In order to better understand and implement, the invention will now be described in detail with reference to the accompanying drawings.
Brief description of the drawings
Fig. 1 is the test strips structure schematic diagram of Respiratory Tract Adenovirus IgA antibody in present invention detection saliva.
Fig. 2 extrudes saliva schematic diagram for syringe.
Fig. 3 is colloidal gold paper slip test 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.
Embodiment
【Embodiment 1】It is prepared by Respiratory Tract Adenovirus IgA antibody diagnostic reagent srip
1st, raw material sources：Goat-anti people IgA antibody is purchased from Sigma companies, and people's IgA antibody thinks biotechnology purchased from Guangzhou promise
Co., Ltd.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 amino acid, entrusts Shanghai gill biochemical corp to be responsible for synthesis, purity is more than altogether
98%, freeze and preserve.
2nd, prepared by goat-anti people IgA antibody colloid gold label pad
In pH value 6.8-7.0 solution, goat-anti people IgA antibody marks to form goat-anti people's IgA antibody glue with colloid gold particle
Body gold label (labelled amount 25ug/ml), confining liquid component are PEG20000 and BSA albumen.Colloid gold label thing is sprayed
Onto glass fibre membrane, dilution parameters 30cm2/ ml, and under the conditions of humidity 15%, 30 DEG C of temperature it is dry 12 it is small when more than,
The colloid gold label pad sealing of preparation is spare.
3rd, nitrocellulose filter is coated with
With coating buffer dilution people 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., will such as attached drawing 1
Both are uniformly sprayed on NC films, and wherein people's IgA antibody is sprayed onto location of C on NC films, Respiratory Tract Adenovirus hexon antigen
Polypeptide fragment is sprayed onto T location on NC films.The NC films sprayed are placed on humidity 10-30%, are dried under the conditions of 20-35 DEG C of temperature
Handle 12 it is small when more than, the sealing of the NC films dried is spare.
4th, assemble, cut
With reference to 1 schematic diagram of attached drawing, by blotting paper 4, nitrocellulose filter 3, colloid gold label pad 2, glass fibre membrane 1 by suitable
Sequence is pasted on pvc backboards 5, is then adjusted cutting machine parameter and is cut, is cut into the test strips of 4mm width.
【Embodiment 2】Sterile absorbent cotton is standby
Purchase the medical absorbent cotton ball of a diameter of 2cm, length is the cotton thread of 8-10cm in binding.The cotton balls handled well is put
Put in pressure cooker, pressure condition 121 DEG C 30 minutes, dry in oven, 100 DEG C of temperature is placed after high pressure, it is dry when the time 4 is small
Cotton balls aluminium foil bag vacuum sealing afterwards.
【Embodiment 3】Saliva sample gathers
The present invention is that sterile absorbent cotton is placed into the sublingual of child, and cotton thread is sucked 3-5 minutes outside mouth.It is such as attached
Fig. 2, cotton balls 22 is placed into syringe 21, then loads onto syringe catch bar, and light and slow promotion, obtains saliva sample 23, uses
2ml tubules preserve.
【Embodiment 4】Sample test
Illustrate with reference to attached drawing 3, Respiratory Tract Adenovirus IgA paper slips are placed into the tubule equipped with saliva sample, sample pad
End in contact saliva sample, is placed 15-30 minutes, then visual results.
, can be with goat-anti people's IgA antibody colloid in label pad if containing Respiratory Tract Adenovirus IgA antibody in saliva sample
Golden label (C lines) combines, the Respiratory Tract Adenovirus hexon antigenic polypeptide fragments detection line (T lines) being up on NC films
Specific bond, forms red stripes (attached drawing 4).If no in sample contain corresponding Respiratory Tract Adenovirus IgA antibody, goat-anti
People's IgA antibody colloid gold label thing can cross detection line (T lines), continue to be up to control line and people's IgA antibody (C lines) combination,
Red stripes (attached drawing 5) are formed on control line.If control line (C lines) does not form red stripes, illustrate that test strips fail,
As a result unreliable (attached drawing 6).
【Embodiment 5】Respiratory Tract Adenovirus IgA colloidal gold diagnosis method of the present invention and fluorescence PCR method Comparative result
Clinical samples 60 are chosen, the saliva sample and oropharyngeal swab specimen of patient are gathered respectively, respectively with respiratory tract adenopathy
Malicious IgA antibody test strip diagnostic method is tested with fluorescence PCR method, as a result such as table 1：
The Respiratory Tract Adenovirus IgA antibody test strip diagnostic method and fluorescence PCR method comparative result of the present invention of table 1
Compared with fluorescence PCR method, sensitivity reaches Respiratory Tract Adenovirus IgA colloidal gold diagnosis methods of the present invention
95.00%, specific 95.00%, the condition of clinical detection can be met.
The invention is not limited in the above embodiment, if the various changes or deformation to the present invention do not depart from the present invention
Spirit and scope, if these changes and deformation belong within the scope of the claim and equivalent technologies of the present invention, then this hair
It is bright to be also intended to comprising these changes and deformation.
Claims (5)
1. a kind of Respiratory Tract Adenovirus IgA antibody test strip, including sample pad, label pad, nitrocellulose filter, water suction
Paper, backboard, it is characterised in that the nitrocellulose filter includes being coated with Respiratory Tract Adenovirus hexon antigen polypeptide
The detection line of fragment and the control line of people's IgA antibody, the amino of the Respiratory Tract Adenovirus hexon antigenic polypeptide fragments
Acid sequence is as described in sequence table Seq ID NO.1.
2. Respiratory Tract Adenovirus IgA antibody test strip according to claim 1, it is characterised in that the sample pad,
Label pad, nitrocellulose filter, blotting paper are adhered on backboard.
3. Respiratory Tract Adenovirus IgA antibody test strip according to claim 2, it is characterised in that the mark
Pad is containing the upper colloidal gold of anti-human IgA antibody mark, and the anti-human IgA antibody is goat-anti people IgA polyclonal antibodies, the anti-human IgA of mouse
Monoclonal antibody, rabbit-anti people IgA monoclonal antibodies or rabbit-anti people's IgA polyclonal antibodies.
4. the Respiratory Tract Adenovirus IgA antibody test strip according to any bar in claim 1-3, it is characterised in that
The ELISA test strip people saliva sample.
5. Respiratory Tract Adenovirus IgA antibody test strip according to claim 4, it is characterised in that the saliva mark
Sterile absorbent cotton by being positioned over the method acquisition that object to be measured is sublingual, is squeezed out again with syringe after absorption saliva by this.
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| CN201610322236.6A CN106168623B (en) | 2016-05-12 | 2016-05-12 | Respiratory Tract Adenovirus IgA antibody test strip and its detection method |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111044728B (en) * | 2019-09-27 | 2023-05-30 | 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 |
| CN111999494B (en) * | 2020-08-18 | 2022-08-26 | 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 |
| CN114527282A (en) * | 2022-02-18 | 2022-05-24 | 嘉兴康源科泰科技发展有限公司 | Detection technology for saliva and nasal cavity double-sample integrated sample adding and application thereof |
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| DE102004041659A1 (en) * | 2004-08-27 | 2006-03-02 | Institut Virion/Serion Gmbh | Test device for the in vitro diagnosis of multi-analyte tests and their use |
| 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 |
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