CN106053803A - Multi-connected detection reagent card adopting immunochromatography and used for respiratory pathogens - Google Patents
Multi-connected detection reagent card adopting immunochromatography and used for respiratory pathogens Download PDFInfo
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- CN106053803A CN106053803A CN201610395679.8A CN201610395679A CN106053803A CN 106053803 A CN106053803 A CN 106053803A CN 201610395679 A CN201610395679 A CN 201610395679A CN 106053803 A CN106053803 A CN 106053803A
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- immunochromatography
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- respiratory pathogen
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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
Abstract
The invention relates to the field of medical examination, in particular to a multi-connected detection reagent card adopting immunochromatography and used for respiratory pathogens. The multi-connected detection reagent card comprises multiple sample pads and multiple nitrocellulose membranes connected with absorbent paper, wherein each sample pad is covered with multiple fluorescent microspheres, and the surface of each fluorescent microsphere is covered with an antibody of one respiratory pathogen; each nitrocellulose membrane connected with the absorbent paper is provided with at least two detection lines, and the detection lines are arranged at the nitrocellulose membrane; each detection line is fixedly covered with an antibody of one respiratory pathogen, and the antibody is in a pairing relation with the antibody, covering the surface of one fluorescent microsphere covering the corresponding sample pad, of one respiratory pathogen. The reagent card is simple, convenient and rapid to operate; different specific test strips can be formed through matching of different nitrocellulose membranes connected with the absorbent paper and the sample pads, different test strips are matched in a mixed manner and put in card paper according to demands, the detection flexibility and the customization property are high, and the detection cost is saved.
Description
Technical field
The present invention relates to field of medical examination, in particular to a kind of multi-joint detection of respiratory pathogen immunochromatography
Reagent card.
Background technology
Respiratory tract infection refers to the respiratory systems such as the nasal cavity of pathogenic infection human body, throat, trachea and bronchus.It is divided into
Respiratory tract infection and lower respiratory infection, upper respiratory tract infection commonly acute upper respiratory tract infection (acute upper
Respir tract infection), refer to the general title of nasal cavity, pharynx or acute throat inflammation, be the modal a kind of biography of respiratory tract
Catch an illness.Commonly encountered diseases is because virus, and minority is caused by antibacterial.Patient is of all ages, sex, professional and regional.Not only have stronger
Infectiousness, and severe complication can be caused.Lower respiratory infection is modal infectious disease, including acute gas
Tracheobronchitis, chronic bronchitis, pneumonia, bronchiectasis etc., by virus, antibacterial, mycoplasma, chlamydia, legion
The microorganisms such as bacterium cause, its preventing and treating should follow put prevention first, Accurate Diagnosis, the principle of in time treatment, must clearly draw during treatment
Act the pathogen infected to select effective antibiotic.
The atypical respiratory pathogens reported has a lot, and modal have legionella pneumophilia, mycoplasma pneumoniae, pneumonia
The hot rickettsia of chlamydia, Q, adenovirus, respiratory syncytial virus, influenza virus, parainfluenza virus etc..Respiratory pathogen because of
The nonspecific symptom of its clinic, major part relies on lab testing method, the germy disease of detection method being applied at present
The cultured tissue cell culture method of substance separation and Culture and virus, serology and direct Detection Method；Indirectly, direct immunofluorescence resists
Body method (IFA/DFA)；Enzyme immunoassay (EIA) (EIA), nucleic acid amplification (PCR).
Although the detection method of existing respiratory pathogens is varied, but it is the most extremely difficult to make a definite diagnosis pathogenic infection.Antibacterial
Pathogen isolation cultivate and cultured tissue cell culture method past of virus is frequently as " goldstandard ", but because its operation is complicated,
The shortcomings such as incubation time length, technical difficulty are big, positive rate is low and be difficulty with, within nearly 10 years, replaced by molecular method.Along with
Developing rapidly of quick diagnosis technology, molecular detection technology heals and becomes perfect, the new mark detected as respiratory pathogen
Standard, but cannot popularize because it requires high to laboratory condition, and the product of commercialization is less, does not forms series, and most sick
Former still can not detect.The extensive application of the development of immunological technique, particularly immunolabelling technique in recent years, as direct, indirectly exempt from
The labelling techniques such as epidemic disease fluorescence method, alkali resistance phosphatase bridging enzyme linked immunosorbent assay so that application immunofluorescence technique detection respiratory pathogens
The method of body is developed and is applied, but this technology yet suffers from numerous shortcoming: unspecific staining problem is not yet fully solved,
The objectivity that result judges is not enough, and detection analytical technology program is complicated, requires the highest to the level professional technology of operator, and
During multiple pathogens joint inspection the longest, complex operation.The most indirectly, chlamydia trachomatis (IFA/DFA) can combine
Become the form of many joint inspections, the immuno-fluorescence assay reagent of the domestic VIRCELL company of import Spain having agency, can examine simultaneously
Surveying 9 respiratory pathogen antibody, in order to clinical assistant diagnosis, but the method there is also complex operation step, and operator are special
Industry level and clinical experience require height, need to use the defects such as accurate operation electron microscope observation.
In view of this, the special proposition present invention.
Summary of the invention
It is an object of the invention to provide a kind of respiratory pathogen immunochromatography multi-link detection reagent card, described
Multi-link detection reagent card is easy and simple to handle quickly, can be used for detecting various respiratory road pathogen simultaneously.
In order to realize the above-mentioned purpose of the present invention, spy by the following technical solutions:
Multiple sample pad, multiple nitrocellulose filter being connected with absorbent paper；
It is coated with multiple fluorescent microsphere in each described sample pad, and every kind of fluorescent microsphere surface is all coated with a kind of breathing
The antibody of road pathogen；
It is provided with at least two detection lines on each nitrocellulose filter being connected with absorbent paper, and described detection line is arranged on
Nitrocellulose filter position；
Fix the antibody being coated with a kind of respiratory pathogen on every detection line, and this antibody wraps in described sample pad
The antibody of the respiratory pathogen of the one of which fluorescent microsphere pan coating of quilt is pair relationhip.
The present invention uses immunochromatography technique, can complete multiple item associations inspection of respiratory pathogen in 15 minutes
Survey, simple and efficient to handle, operator's level professional technology is required low, is suitable for the other detection of clinical bed, quickly obtains detection
Result.
The quantity of usual described sample pad is identical with the described nitrocellulose filter being connected with absorbent paper；
In sample pad, the kind of the antibody on coated fluorescent microsphere can be the same or different；
In use, sample pad is fitted together available reagent paper with the corresponding nitrocellulose filter being connected with absorbent paper
Bar.
Preferably, respiratory pathogen immunochromatography multi-link detection reagent card as above, described sample pad and described
The side of the nitrocellulose filter being connected with absorbent paper is additionally provided with coated film and end liner.
Wherein, institute's coated film be positioned in the middle of described sample pad and described end liner and described in be connected with the nitric acid of absorbent paper fine
In the middle of dimension element film and described end liner.
Preferably, respiratory pathogen immunochromatography multi-link detection reagent card as above, described multi-link detection reagent
Card also includes getting stuck；Described get stuck on be provided with sample pipetting volume district and reagent paper and place screens.
Described paper is placed screens and is i.e. assembled the draw-in groove of sample pad and the corresponding nitrocellulose filter being connected with absorbent paper.
It is further preferred that respiratory pathogen immunochromatography multi-link detection reagent card as above, described reagent paper is put
Put and be provided with interface channel between sample adding area end and the sample application zone of screens.
It is further preferred that respiratory pathogen immunochromatography multi-link detection reagent card as above, described reagent paper is put
Putting card bit quantity is 2～4.
Preferably, respiratory pathogen immunochromatography multi-link detection reagent card as above, described sample pad is coated
Have 2～17 kind of fluorescent microsphere.
Preferably, respiratory pathogen immunochromatography multi-link detection reagent card as above, described detection line is 2～5
Bar.
It is further preferred that respiratory pathogen immunochromatography multi-link detection reagent card as above, the most adjacent two
Detection line is parallel to each other, and is spaced apart 3～8mm.
Preferably, respiratory pathogen immunochromatography multi-link detection reagent card as above, it is characterised in that described company
Having on the nitrocellulose filter of absorbent paper and be provided with control line, on described control line, fixed packet is had described fluorescent microsphere surface to wrap
The two of the antibody of the respiratory pathogen of quilt resist.
Arrange the purpose of control line be in order to avoid or reduce the appearance of false negative result.
Preferably, respiratory pathogen immunochromatography multi-link detection reagent card as above, it is characterised in that described many
Link detection reagent card also includes fluorescence detector.
Described fluorescence detector is used for reading the fluorescence signal on detection line, it is achieved the automatic interpretation to result, reduces main
See interpretation.
Preferably, respiratory pathogen immunochromatography multi-link detection reagent card as above, described respiratory pathogen
Including influenza A virus, Influenza B virus, respiratory syncytial virus, Respiratory Tract Adenovirus, parainfluenza virus, rhinovirus,
Echovirus, Measles virus, rubella virus, adenovirus, hemophilus influenza, streptococcus pneumoniae, staphylococcus, legionella pneumophilia,
The hot rickettsia of mycoplasma pneumoniae, Chlamydia pneumoniae, Q.
Compared with prior art, the invention have the benefit that
1), the present invention use immunochromatography technique, can complete in 15 minutes respiratory pathogen multiple projects connection
Close detection, simple and efficient to handle, operator's level professional technology is required low, is suitable for the other detection of clinical bed, quickly obtains
Testing result.
2), different specifically trying can be formed by the different nitrocellulose filters being connected with absorbent paper from sample pad collocation
Paper slip, strong further according to needing different test strips mashed up loading paperboards, the motility of detection and customization, it is beneficial to targetedly
To in the face of different detection case, save detection time and testing cost.
3), the present invention provide multi-link detection reagent card also include supporting fluorescence detector, can realize to result from
Dynamic interpretation, reduces the error that the supervisor that operator brings judges, and the advantage such as also can save historical data.
Accompanying drawing explanation
In order to be illustrated more clearly that the specific embodiment of the invention or technical scheme of the prior art, below will be to specifically
In embodiment or description of the prior art, the required accompanying drawing used is briefly described, it should be apparent that, in describing below
Accompanying drawing is some embodiments of the present invention, for those of ordinary skill in the art, before not paying creative work
Put, it is also possible to obtain other accompanying drawing according to these accompanying drawings.
Fig. 1 is the influenza A virus in the embodiment of the present invention 1 and Influenza B virus combined detection test paper；
Fig. 2 is the respiratory syncytial virus in the embodiment of the present invention 2, Respiratory Tract Adenovirus and parainfluenza virus joint-detection
Test strips；
Fig. 3 is respiratory pathogen immunochromatography multi-link detection reagent card in the embodiment of the present invention 3.
Reference:
101 nitrocellulose filters being connected with absorbent paper；
102 absorbent paper；
103 nitrocellulose filters；
104 sample pad；
105A first detects line；
105B second detects line；
106 influenza A viruss and Influenza B virus combined detection test paper；
107 control lines；
201 nitrocellulose filters being connected with absorbent paper；
202 absorbent paper；
203 nitrocellulose filters；
204 sample pad；
205A first detects line；
205B second detects line；
205C second detects line；
206 respiratory syncytial virus, Respiratory Tract Adenovirus and parainfluenza virus combined detection test paper；
207 control lines；
301 get stuck；
302 sample application zone；
303 interface channels.
Detailed description of the invention
Below in conjunction with embodiment, embodiment of the present invention are described in detail, but those skilled in the art will
Understanding, the following example is merely to illustrate the present invention, and is not construed as limiting the scope of the present invention.In embodiment unreceipted specifically
Condition person, the condition advised according to normal condition or manufacturer is carried out.Agents useful for same or instrument unreceipted production firm person, be
Can be by the commercially available conventional products bought and obtain.
Embodiment 1
Present embodiments provide influenza A virus and the assemble method of Influenza B virus combined detection test paper, such as figure
Shown in 1:
The nitrocellulose filter 101 being connected with absorbent paper is divided into absorbent paper 102 and nitrocellulose filter 103 two parts, by sample
One end combination of product pad 104 and nitrocellulose filter 103 links together and obtains influenza A virus and Influenza B virus connection
Close test strip 106.
The side detecting line dorsad of described sample pad 104 and the described nitrocellulose filter 101 being connected with absorbent paper also sets
It is equipped with coated film and end liner.
Wherein it is provided with the first detection line 105A and second detection line 105B on nitrocellulose filter 103；
Described first detection line 105A and the second detection line 105B be arranged in parallel, and is spaced apart 8mm.
It is coated fluorescent microsphere in sample pad 104, each fluorescent microsphere surface is marked with the first of influenza A virus and resists
One in the first antibody (rabbit source) of body (mice source) and Influenza B virus；
The fixing second antibody being coated with influenza A virus on described first detection line 105A, on the second detection line 105B
It is coated with the second antibody of Influenza B virus；
The first antibody of described influenza A virus be combined with the second antibody of described influenza A virus for same first
Type influenza virus surface antigens, and it is monoclonal antibody, difference is that the epitope combined is different, i.e. both antibody is
Pairing antibody.
Same, the first antibody of described Influenza B virus with the second antibody of described Influenza B virus is combined is
Same Influenza B virus surface antigen, and it is monoclonal antibody, difference is that the epitope combined is different, i.e. both
Antibody is pairing antibody.
Wherein it is additionally provided with control line 107 on nitrocellulose filter 103；
On described control line 107, fixing the two of the goat anti-mouse that is coated with resists.As long as two on control line are anti-also has work
Property, then illustrating that the preservation condition of this strip is good, remaining activated feedstock all keeps activity good, it is possible to ensure the effective of strip
Property.So control line can only arrange one.
Embodiment 2
Present embodiments provide respiratory syncytial virus, Respiratory Tract Adenovirus and parainfluenza virus combined detection test paper
Assemble method, as shown in Figure 2:
The nitrocellulose filter 201 being connected with absorbent paper is divided into absorbent paper 202 and nitrocellulose filter 203 two parts, by sample
One end combination of product pad 204 and nitrocellulose filter 203 link together obtain respiratory syncytial virus, Respiratory Tract Adenovirus and
Parainfluenza virus combined detection test paper 206.
The side detecting line dorsad of described sample pad 204 and the described nitrocellulose filter 201 being connected with absorbent paper also sets
It is equipped with coated film and end liner.
Wherein it is provided with the first detection line 205A, the second detection line 205B and the 3rd detection line on nitrocellulose filter 203
205C；
Described first detection line 205A, the second detection line 205B and the 3rd detection line 205C be arranged in parallel, and adjacent two
That detects line is spaced apart 5mm.
It is coated fluorescent microsphere in sample pad 204, each fluorescent microsphere surface is marked with respiratory syncytial virus, respiratory tract
One in the first antibody of adenovirus and parainfluenza virus；And these antibody are mice source.
The fixing second antibody being coated with respiratory syncytial virus on described first detection line 205A, the second detection line 205B
On be coated with the second antibody of Respiratory Tract Adenovirus, the 3rd detection line 205C is coated with the second antibody of parainfluenza virus,；
The first antibody of described respiratory syncytial virus be combined with the second antibody of described influenza A virus for same
Respiratory syncytial virus proteantigen, and it is monoclonal antibody, difference is that the epitope combined is different, i.e. both resists
Body is pairing antibody.
Same, the first antibody of described Respiratory Tract Adenovirus and parainfluenza virus and described Respiratory Tract Adenovirus and sidestream
The second antibody of Influenza Virus is also for pairing antibody.
Wherein it is additionally provided with control line 207 on nitrocellulose filter 203；
On control line 207, fixing the two of the goat anti-mouse that is coated with resists.
Embodiment 3
Present embodiments provide assembling and the using method of a kind of respiratory pathogen immunochromatography multi-link detection reagent card,
Described respiratory pathogen immunochromatography multi-link detection reagent card can be used for influenza A virus, Influenza B virus, respiratory tract
Syncytial virus, Respiratory Tract Adenovirus and the joint-detection of 5 projects of parainfluenza virus.Specifically refer to shown in Fig. 3:
Get stuck and there is on 301 reagent paper place screens, sample application zone 302 and interface channel 303；Preferably, described interface channel
303 is hydrophilic pathway, connects reagent paper and places screens and sample application zone 302.
It should be noted that the shape of sample application zone 302 may be configured as the ellipse of diagram, it is possible to be set to strip or square
The shapes such as shape.
Preferably, interface channel 303 can also be provided at get stuck 301 inside, only open rear end and connect sample-adding respectively for two
Screens placed by district 302 and reagent paper.
By the influenza A virus assembled in embodiment 1 and Influenza B virus combined detection test paper 106 and embodiment 2
The respiratory syncytial virus of middle assembling, Respiratory Tract Adenovirus and parainfluenza virus combined detection test paper 206 load and get stuck 301
Reagent paper is placed in screens, and makes sample pad 104 and sample pad 204 be positioned at sample application zone 302 near-end, absorbent paper 102 and water suction
Paper 202 is positioned at sample application zone 302 far-end.
Testing sample is added dropwise to sample application zone 302, and under chromatography effect, sample flows to influenza A through interface channel 303
Virus and Influenza B virus combined detection test paper 106 and respiratory syncytial virus, Respiratory Tract Adenovirus and parainfluenza virus
Combined detection test paper 206, and flow through the sample pad of two test strips and the detection line in nitrocellulose filter district successively.
If containing determinand in sample, then can combine by its corresponding first antibody when flowing through sample pad, form fluorescence
Microsphere-first antibody combination-antigenic compound, described fluorescent microsphere-first antibody combination-antigenic compound is along with sample solution
Continue towards nitrocellulose filter district, and fix coated corresponding second antibody on the detection line in nitrocellulose filter district
In conjunction with, and then be fixed on corresponding detection line.
After 15 minutes, reagent card is inserted supporting Immunofluorescence test and can be automatically obtained the judgement to result.
Embodiment 4
Based on embodiment 2, the sample pad in embodiment 2 and detection line are replaced.
It is coated fluorescent microsphere in sample pad, each fluorescent microsphere surface is marked with mycoplasma pneumoniae, Chlamydia pneumoniae, Q
One in hot rickettsia, legionella pneumophilia, staphylococcic first antibody；
Detection line is five, is followed successively by:
First detection line, the second detection line, the 3rd detection line, the 4th detection line and the 5th detection line, on five detection lines
Corresponding fixing coated antibody is mycoplasma pneumoniae, Chlamydia pneumoniae, the hot rickettsia of Q, legionella pneumophilia, staphylococcic
Second antibody, and the second antibody of corresponding pathogen and first antibody are for matching antibody.
Other embodiments with embodiment 2, thus obtain mycoplasma pneumoniae, Chlamydia pneumoniae, the hot rickettsia of Q, addicted to lung
Legionella, staphylococcus combined detection test paper.
Embodiment 5
Based on embodiment 2, the sample pad in embodiment 2 and detection line are replaced.
It is coated fluorescent microsphere in sample pad, each fluorescent microsphere surface is marked with rhinovirus, Measles virus, rubella
Poison, echovirus first antibody in one；
Detection line is four, is followed successively by:
First detection line, the second detection line, the 3rd detection line and the 4th detection line, fixed packet corresponding on five detection lines
The antibody of quilt is the second antibody of rhinovirus, Measles virus, rubella virus, echovirus, and the second antibody of corresponding pathogen
It is pairing antibody with first antibody.
Other embodiments, with embodiment 2, thus obtain rhinovirus, Measles virus, rubella virus, echovirus combine inspection
Test paper slip.
Embodiment 6
Reequiping based on embodiment 3, method of modifying is, chooses and has getting stuck of 4 reagent paper placement screens, will be real
Executing the test strips assembled in example 1, embodiment 2, embodiment 4, embodiment 5 and put into wherein, other set-up modes are with embodiment 3, i.e.
Obtain the reagent card that can simultaneously respiratory pathogen in 14 be detected.
Last it is noted that various embodiments above is only in order to illustrate technical scheme, it is not intended to limit；To the greatest extent
The present invention has been described in detail by pipe with reference to foregoing embodiments, but it will be understood by those within the art that: its
Still the technical scheme described in foregoing embodiments can be modified, or to the most some or all of technical characteristic
Carry out equivalent；And these amendments or replacement, do not make the essence of appropriate technical solution depart from various embodiments of the present invention skill
The scope of art scheme.
Claims (10)
1. a respiratory pathogen immunochromatography multi-link detection reagent card, it is characterised in that including:
Multiple sample pad, multiple nitrocellulose filter being connected with absorbent paper；
It is coated with multiple fluorescent microsphere in each described sample pad, and every kind of fluorescent microsphere surface is all coated with a kind of respiratory tract disease
The antibody of substance；
It is provided with at least two detection lines on each nitrocellulose filter being connected with absorbent paper, and described detection line is arranged on nitric acid
Cellulose membrane position；
The fixing antibody being coated with a kind of respiratory pathogen on every detection line, and this antibody is coated with in described sample pad
The antibody of the respiratory pathogen of one of which fluorescent microsphere pan coating is pair relationhip.
2. respiratory pathogen immunochromatography multi-link detection reagent card as claimed in claim 1, it is characterised in that described sample
The side of pad and the described nitrocellulose filter being connected with absorbent paper is additionally provided with coated film and end liner.
3. respiratory pathogen immunochromatography multi-link detection reagent card as claimed in claim 1, it is characterised in that described multi-joint
Detectable card also includes getting stuck；Described get stuck on be provided with sample pipetting volume district and reagent paper and place screens.
4. respiratory pathogen immunochromatography multi-link detection reagent card as claimed in claim 3, it is characterised in that described reagent paper
Place and be provided with interface channel between sample adding area end and the sample application zone of screens.
5. respiratory pathogen immunochromatography multi-link detection reagent card as claimed in claim 4, it is characterised in that described reagent paper
Placing card bit quantity is 2～4.
6. respiratory pathogen immunochromatography multi-link detection reagent card as claimed in claim 1, it is characterised in that described sample
2～17 kind of fluorescent microsphere it are coated with on pad.
7. respiratory pathogen immunochromatography multi-link detection reagent card as claimed in claim 1, it is characterised in that described detection
Line is 2～5.
8. respiratory pathogen immunochromatography multi-link detection reagent card as claimed in claim 1, it is characterised in that described in be connected with
Being provided with control line on the nitrocellulose filter of absorbent paper, on described control line, fixed packet is had described fluorescent microsphere pan coating
Respiratory pathogen antibody two resist.
9. respiratory pathogen immunochromatography multi-link detection reagent card as claimed in claim 1, it is characterised in that described multi-joint
Detectable card also includes fluorescence detector.
10. the respiratory pathogen immunochromatography multi-link detection reagent card as described in any one of claim 1～9, its feature exists
Influenza A virus is included in, described respiratory pathogen, Influenza B virus, respiratory syncytial virus, Respiratory Tract Adenovirus,
Parainfluenza virus, rhinovirus, echovirus, Measles virus, rubella virus, adenovirus, hemophilus influenza, streptococcus pneumoniae, Portugal
Grape coccus, legionella pneumophilia, mycoplasma pneumoniae, Chlamydia pneumoniae, the hot rickettsia of Q.
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