CN204495834U - Detect the test strips of chicken trachitis virus - Google Patents
Detect the test strips of chicken trachitis virus Download PDFInfo
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- CN204495834U CN204495834U CN201520097970.8U CN201520097970U CN204495834U CN 204495834 U CN204495834 U CN 204495834U CN 201520097970 U CN201520097970 U CN 201520097970U CN 204495834 U CN204495834 U CN 204495834U
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Abstract
The utility model belongs to a kind of utensil detecting chicken trachitis virus, particularly relate to a kind of test strips detecting chicken trachitis virus, comprise: detection layers, it comprises the sample adsorption layer, binding layer, cellulose rete and the water accepting layer that connect successively, and described binding layer includes the first antibody of the chicken trachitis virus of colloid gold label; Detection line, it is arranged on described cellulose rete, and described detection line comprises the second antibody of the chicken trachitis virus of bag quilt, and described first antibody is different with the site that the scorching virus of described second antibody and described chicken trunnion combines; Control line, it is arranged on described cellulose rete, and described control line comprises the sheep/rabbit anti-mouse immunoglobulin G of bag quilt.The utility model provides a kind of result intuitive display, accurately, detects the test strips of ILTV fast, and compared with other detection methods, fast, testing cost is cheap for this test strips high specificity, highly sensitive, easy and simple to handle, result display.
Description
Technical field
The utility model belongs to a kind of utensil detecting chicken trachitis virus, particularly relates to a kind of test strips detecting chicken trachitis virus.
Background technology
Infectious laryngotracheitis of chicken (Infectious laryngotracheitis, ILT) be by avian infectious laryngotracheitis virus (Infectious laryngotracheitis virus, ILTV) chicken caused is acute, hot, high degree in contact upper respiratory infectious disease, infected chicken shows as expiratory dyspnea, expectoration band blood mucus, tracheal mucosa oedema and hemorrhage etc., and cause infected chicken death and laying hen egg production to decline, cause serious economic loss to poultry husbandry.
ILT is widely current in all over the world, has a strong impact on aviculture and develops in a healthy way.China finds this disease the fifties in last century, is now widely current, and is one of important epidemic disease of harm poultry husbandry.Due to ILTV belong to herpesviral, can latent infection in trigeminal neuralgia cell, avoid host immune to remove, thus in chicken group sustainable existence.And, be subject to stress latent infection chicken, ILTV can be activated, and massive duplication and discharge.Therefore, chicken group, once infect ILTV, just cannot remove, thus cause irregular morbidity, the extreme influence sound development of China's poultry husbandry.
In time, quick diagnosis ILT, be the prerequisite of effectively control ILT.The diagnosis of current ILT mainly contains Routine Test Lab and detects and the large class methods of molecular biology two.Conventional method mainly contains the Serologic detection of Virus Isolation and ILTV antibody, comprises the methods such as agar immunodiffusion test (AGP), indirect immuno-fluorescence assay (IFA), ELISA.Molecular biology method mainly contains PCR and Nucleic Acid Probe Technique etc.
The Viral isolation of ILTV, the detection technique complicated operation such as AGP, IFA, ELISA, PCR and nucleic acid probe, the instrument and equipment that needs are expensive, process are longer, time and effort consuming, need particular instrument equipment and professional and technical personnel's operation, be difficult to popularize in basic unit, the quick diagnosis needs at production line or epidemic disease scene can not be met.Therefore, detection technique that a kind of quick detection fowl avian infectious laryngotracheitis virus infects is studied extremely important and urgent.
Utility model content
An object of the present utility model solves at least the problems referred to above or defect, and provide the advantage will illustrated at least below.
The utility model also has an object to there is provided a kind of test strips detecting chicken trachitis virus, can detect chicken trachitis virus directly perceived, accurate, fast.
Another object of the utility model is, provides a kind of test strips detecting chicken trachitis virus, and compared with other detection methods, fast, and testing cost is cheap for the display of this test strips high specificity, highly sensitive, easy and simple to handle, result.
In order to realize, according to these objects of the present utility model and other advantage, providing a kind of test strips detecting chicken trachitis virus, comprising:
Detection layers, it comprises the sample adsorption layer, binding layer, cellulose rete and the water accepting layer that connect successively, described sample adsorption layer defines the first end for contacting detected sample, described water accepting layer defines the second end away from detected sample, and described binding layer includes the first antibody of the chicken trachitis virus of colloid gold label;
Detection line, it is arranged on described cellulose rete, described detection line comprises the second antibody of the chicken trachitis virus of bag quilt, described first antibody is different with the site that the scorching virus of described second antibody and described chicken trunnion combines, and described first antibody and described chicken trunnion scorching virus combines, do not affect described second antibody and combine with described chicken trunnion inflammation is viral;
Control line, it is also arranged on described cellulose rete and than described detection line away from described first end, described control line comprises the sheep/rabbit anti-mouse immunoglobulin G of bag quilt.
Preferably, wherein, described first antibody is monoclonal antibody or the polyclonal antibody of chicken trachitis virus, and described second antibody is polyclonal antibody or the monoclonal antibody of chicken trachitis virus.
Preferably, wherein, also comprise:
Supporting layer, described detection layers is fixed on the upper surface of described supporting layer.
Preferably, wherein, also comprise:
First diaphragm, it is arranged on above described detection layers, and one end of described first diaphragm bends downwards and envelopes described first end, and the other end extends to the top of described cellulose rete one end;
Second diaphragm, it is arranged on the top of described detection layers, and one end of described second diaphragm bends downwards and envelopes described second end, and the other end extends to the top of the described cellulose rete other end;
Wherein, described first diaphragm is white, and the color of described second diaphragm is not identical with the color of described first diaphragm.
Preferably, wherein, described first diaphragm is provided with sample mark line, and described sample mark line is 0.3 ~ 0.7cm place with the distance away from the side of the described sample adsorption layer of described first end.
Preferably, wherein, described cellulose rete is nitrocellulose filter, any one in cellulose membrane, carboxylated cellulose film and PVDF membrane make.
Preferably, wherein, described sample adsorption layer is made up of any one in glass wool, nylon fiber and dacron.
Preferably, wherein, described binding layer is made up of any one in glass wool, nylon fiber and dacron.
Preferably, wherein, described supporting layer is made up of the hard plastic sheet not absorbing water or cardboard bar; Described water accepting layer thieving paper is made.
Preferably, wherein, the spread pattern of described detection line and described control line be " || ", " ++ ",
with
in any one.
The beneficial effects of the utility model
1, the utility model is according to the ultimate principle of ELISA, virus is detected with immune chromatography test paper, the diafiltration of miillpore filter is utilized to concentrate and capillarity, antigen-antibody reaction is forwarded on solid phase filter membrane by the Traditional liquid phase environment of ELISA and carries out fast, and adopt collaurum trace to replace enzyme trace, with the colour developing situation of direct visual perception collaurum, immediately obtain testing result, more easier, quick than serological methods such as ELISA.
2, the test strips of detection chicken trachitis virus that provides of the utility model, detection specificity is strong, and susceptibility is high, the albumen of the corresponding virus of 2 nanograms can be detected.
3, the test strips of detection chicken trachitis virus that provides of the utility model, easy and simple to handle, fast.Without the need to additional any Other Instruments and reagent when using this test strips, as long as its test lead to be inserted in measuring samples liquid about 30 seconds, then testing result can be judged at about 5 minutes.
4, the test strips of detection chicken trachitis virus that provides of the utility model, result intuitive display, accurately, simple and clear, not easily there is false negative and false positive erroneous judgement.
5, the test strips of detection chicken trachitis virus that provides of the utility model, reduces investment and testing cost.
6, the test strips of detection chicken trachitis virus that provides of the utility model, its applied range, is convenient to popularity application; This test strips simple to operate, becomes " single step " or " foolproof ", and is convenient for carrying and preserves, can meet the needs of different levels personnel, have wide market outlook and good economical, societal benefits.
Accompanying drawing explanation
Fig. 1 is the plan structure schematic diagram of the test strips of detection chicken trachitis virus described in the utility model;
Fig. 2 is the side structure schematic diagram of the test strips of detection chicken trachitis virus described in the utility model.
Embodiment
Below in conjunction with accompanying drawing, the utility model is described in further detail, can implements according to this with reference to instructions word to make those skilled in the art.
Should be appreciated that used hereinly such as " to have ", one or more other element do not got rid of in " comprising " and " comprising " term or the existence of its combination or interpolation.
Detect a test strips for chicken trachitis virus, comprise supporting layer 1; The sample adsorption layer 2 connected successively, binding layer 3, cellulose rete 4 and water accepting layer 5; Detection line 6; Control line 7; First diaphragm 8-1; Second diaphragm 8-2; Sample mark line 9;
Sheep/rabbit anti-mouse the immunoglobulin G (IgG) of bag quilt is comprised for control line, and the polyclonal antibody of chicken trachitis virus (ILTV) or the monoclonal antibody of bag quilt is comprised for detection line, and it is as follows to include the preparation method of chicken trachitis virus (ILTV) monoclonal antibody of colloid gold label or polyclonal antibody for binding layer:
The preparation of 1, sheep/rabbit anti-mouse IgG or sheep/rabbit anti-rabbit/sheep IgG
Extract the IgG in mice serum with saturated ammonium sulfate method: get 1 part of serum and add 2 parts of PBS liquid (pH7.2) mixings, add the mixing of equal-volume saturated ammonium sulfate liquid, to put in 4 DEG C of refrigerators 2 hours, at 4 DEG C, the centrifugal 15min of 10000r/min, abandon supernatant; With appropriate PBS liquid (pH7.2) dissolution precipitation, adding saturated ammonium sulfate liquid to its ultimate density is 33%, to put in 4 DEG C of refrigerators 2 hours, at 4 DEG C, centrifugal 15min under 10000r/min condition, abandon supernatant, with a small amount of PBS liquid (pH7.2) dissolution precipitation, to put in 4 DEG C of refrigerators with PBS liquid (pH7.2) dialyzed overnight, change liquid 2-3 time, at 4 DEG C, centrifugal 15min under 10000r/min condition, collect supernatant, measure its protein concentration with ultraviolet spectrophotometer.With 50 μ g ~ 100 μ g (IgG)/kg body weight through subcutaneous or intramuscular injection negative antibody Healthy Sheep or rabbit 3-4 time, final immunization is after 20 days, and venous blood collection, measures its serum antibody titer 1 with ELISA
:more than 2000, Culling heart blood or arteria carotis bloodletting, collect its hyper-immune serum, with the IgG of saturated ammonium sulfate method extraction sheep (rabbit) anti-mouse/chicken, (its extracting method is identical with said extracted mice serum IgG, no longer repeat), in like manner prepare sheep/rabbit anti-rabbit/sheep IgG, for marking the contrast trace of the utility model test strips.
2, the preparation of anti-ILTV monoclonal antibody
Differential centrifugation, sucrose density gradient centrifugation concentrate ILTV CEF embryo toxicity, purifying, prepare immunogene after formalin-inactivated with freund adjuvant emulsification.
With the immunogen immune Balb/c system mouse three times of 50-100 μ g/ dosage, every minor tick 15-30 days; 3-4 days after third time booster immunization, by the bloodletting of immune mouse eyeball, draws neck lethal, in 75% alcohol-pickled 5-10min, asepticly gets its splenocyte; Shred and through 100 order nylon net filters, with GNK washing lotion suspendible splenocyte, the centrifugal 10min of 1000r/min, collects splenocyte; By 1 × 10
8individual splenocyte and 2 × 10
7-5 × 10
7individual NS0 myeloma cell's mixing, after using GNK washing lotion suspendible again, 1000r/min is centrifugal, and 10min abandons supernatant, PEG-1500 (pH8.5-pH 9.0) the limit edged that cell precipitation slowly adds 0.7-1.0mL in the water-bath of 37 DEG C in 1min shakes, then GNK washing lotion 15ml is slowly added, to stop the effect of PEG; 37 DEG C of water-bath 5min, 1000r/min is centrifugal, and 10min abandons supernatant, is resuspended in by cell precipitation in 1640/HAT Selective agar medium, and adds 96 well culture plates (200 μ L/ hole), is placed in 37 DEG C, 5%CO
2after cultivating 7-10 days in incubator, with the purifying ILTV gB albumen bag of 5-10 μ g/mL by 96 hole ELISA Plate, detect the culture supernatant of hybridoma with enzyme linked immunosorbent assay (ELISA), picking strong positive cell clone (OD
450>=0.5), the limiting dilution assay of continuous three times carries out subclone, and finally screening obtains the monoclonal cell of the anti-ILTV of specificity.
This hybridoma chromosome number is 92-98, and the monoclonal antibody of the anti-ILTV of its secretion is reacted with ILTV specifically, and affinity constant reaches 10
9-10, light chain subtype is κ or λ, and heavy chain subgroup is IgG
1, IgG
2a, IgG
2b, IgG
3, for the preparation of gold mark monoclonal antibody.
3, the preparation of anti-ILTV gold mark monoclonal antibody and gold mark monoclonal antibody tunica fibrosa
Expanded by the monoclonal cell of the anti-ILTV of specificity screened and cultivate, PBS washes the centrifugal 10min of rear 1000r/min, and collecting cell, with 1 × 10
6-2 × 10
6individual/only to carry out lumbar injection to the female mouse of multiparity, gather mouse ascites after 10-20 days, the supernatant after the centrifugal 10min of 4000r/min is required monoclonal antibody ascites.
With reduction of sodium citrate legal system for aurosol: the 0.5-2% citric acid three sodium solution adding 2-4mL in the 0.01-0.05% aqueous solution of chloraurate of 50-100mL boiling, after reaction, obtain the collaurum of diameter about 15nm.With the K of 0.1mol/L
2cO
3adjust collaurum pH value to 8.5-9.5, trace with 1: 1000-1300 is than treating that the anti-ILTV monoclonal antibody ascites of trace adds in the aurosol of pH8.5-9.5, after trace 10min, PEG-10000 to the PEG-10000 final concentration adding 20wt% reaches 0.05%, at 4 DEG C, the centrifugal 20min of 1500-3000r/min, remove unconjugated colloid gold particle, at 4 DEG C, centrifugal 1 hour of 15000r/min, abandon supernatant, after obtaining the golden labeling antibody protein mixture of preliminary purification, with propylene glucosan S-400 column chromatography, separation and purification gold mark albumen, obtain the anti-ILTV monoclonal antibody of collaurum trace.
The said monoclonal antibody of 1: 100-1: the 500 collaurum traces diluted is adsorbed in processed glass cotton (nylon fiber or dacron), low-temperature vacuum drying at 4 DEG C, i.e. obtained anti-ILTV gold mark monoclonal antibody tunica fibrosa.
4, the preparation of anti-ILTV polyclonal antibody
The ILTV embryo toxicity of the same use deactivation, separately repeatedly immunity inoculation negative antibody Healthy Sheep or rabbit.Final immunization posterior vein blood sampling in 20 days, measure its serum antibody titer more than 1: 1024 with ELISA, Culling heart blood or arteria carotis bloodletting, collect its hyper-immune serum, the IgG antibody (method is identical with extraction mice serum IgG, does not repeat) in serum is extracted with saturated ammonium sulfate method.
Embodiment 1
A kind of test strips detecting chicken trachitis virus comprises:
Supporting layer 1, it is made with plastic slice bar;
Detection layers, it comprises the sample adsorption layer 2 connected successively, its fibrage made with glass wool; Binding layer 3, it is the fibrage that the glass wool of the anti-ILTV monoclonal antibody being attached with colloid gold label is made; Cellulose rete 4, its cellulose rete made for nitrocellulose filter; Water accepting layer 5, it is the water accepting layer be made up of absorbent filter;
Detection line 6; It is on cellulose rete 4, detects trace 6 (code name T) with anti-ILTV polyclonal antibody IgG solution mark;
Control line 7; It is on cellulose rete 4, with sheep (rabbit) anti-mouse IgG solution mark contrast trace 7 (code name C);
First diaphragm 8-1; It is the white diaphragm of the test lead covered above sample adsorption fibrage 2 and binding layer 3,
Second diaphragm 8-2; Water accepting layer 5 is coated with yellow color diaphragm 8-2.
Sample mark line 9; 0.5cm place, sample adsorption fibrage 2 side is partial to by the first diaphragm that the intersection of sample adsorption layer 2 and binding layer 3 is corresponding, and the right-hand member of sample mark line is printed on arrow and MAX printed words,
Wherein, sample adsorption layer 2, binding layer 3, cellulose rete 4, each layer of water accepting layer 5 are pasted onto successively on supporting layer 1, the intersection fiber spliced each other crosses one another infiltration; The spread pattern of detection line and control line is " || ".
Embodiment 2
A kind of test strips detecting chicken trachitis virus comprises:
Supporting layer 1, it is made with plastic slice bar;
Detection layers, it comprises the sample adsorption layer 2 connected successively, its fibrage made with glass wool; Binding layer 3, it is the fibrage that the nylon fiber of the anti-ILTV polyclonal antibody being attached with colloid gold label is made; Cellulose rete 4, its cellulose rete made for nitrocellulose filter; Water accepting layer 5, it is the water accepting layer be made up of absorbent filter;
Detection line 6; It is on cellulose rete 4, detects trace 6 (code name T) with anti-ILTV monoclonal antibody IgG solution mark;
Control line 7; It is on cellulose rete 4, with sheep (rabbit) anti-rabbit (sheep) IgG solution mark contrast trace 7 (code name C);
First diaphragm 8-1; It is the white diaphragm of the test lead covered above sample adsorption fibrage 2 and binding layer 3,
Second diaphragm 8-2; Water accepting layer 5 is coated with red color diaphragm 8-2.
Sample mark line 9; 0.3cm place, sample adsorption fibrage 2 side is partial to by the first diaphragm that the intersection of sample adsorption layer 2 and binding layer 3 is corresponding, and the right-hand member of sample mark line is printed on arrow and MAX printed words,
Wherein, sample adsorption layer 2, binding layer 3, cellulose rete 4, each layer of water accepting layer 5 are pasted onto successively on supporting layer 1, the intersection fiber spliced each other crosses one another infiltration; The spread pattern of detection line and control line is " ++ ".
Embodiment 3
A kind of test strips detecting chicken trachitis virus comprises:
Supporting layer 1, it is made with plastic slice bar;
Detection layers, it comprises the sample adsorption layer 2 connected successively, its fibrage made with glass wool; Binding layer 3, it is the fibrage that the dacron of the anti-ILTV monoclonal antibody being attached with colloid gold label is made; Cellulose rete 4, its cellulose rete made for polyvinylidene fluoride; Water accepting layer 5, it is the water accepting layer be made up of absorbent filter;
Detection line 6; It is on cellulose rete 4, detects trace 6 (code name T) with anti-ILTV polyclonal antibody IgG solution mark;
Control line 7; It is on cellulose rete 4, with sheep (rabbit) anti-mouse IgG solution mark contrast trace 7 (code name C);
First diaphragm 8-1; It is the white diaphragm of the test lead covered above sample adsorption fibrage 2 and binding layer 3,
Second diaphragm 8-2; Water accepting layer 5 is coated with blue color diaphragm 8-2.
Sample mark line 9; 0.7cm place, sample adsorption fibrage 2 side is partial to by the first diaphragm that the intersection of sample adsorption layer 2 and binding layer 3 is corresponding, and the right-hand member of sample mark line is printed on arrow and MAX printed words,
Wherein, sample adsorption layer 2, binding layer 3, cellulose rete 4, each layer of water accepting layer 5 are pasted onto successively on supporting layer 1, the intersection fiber spliced each other crosses one another infiltration; The spread pattern of detection line and control line is " // "., and
in any one
Embodiment 4
A kind of test strips detecting chicken trachitis virus comprises:
Supporting layer 1, it is made with plastic slice bar;
Detection layers, it comprises the sample adsorption layer 2 connected successively, its fibrage made with glass wool; Binding layer 3, it is the fibrage that the nylon fiber of the anti-ILTV polyclonal antibody being attached with colloid gold label is made; Cellulose rete 4, its cellulose rete made for polyvinylidene fluoride; Water accepting layer 5, it is the water accepting layer be made up of absorbent filter;
Detection line 6; It is on cellulose rete 4, detects trace 6 (code name T) with anti-ILTV monoclonal antibody IgG solution mark;
Control line 7; It is on cellulose rete 4, with sheep (rabbit) anti-rabbit (sheep) IgG solution mark contrast trace 7 (code name C);
First diaphragm 8-1; It is the white diaphragm of the test lead covered above sample adsorption fibrage 2 and binding layer 3,
Second diaphragm 8-2; Water accepting layer 5 is coated with green color diaphragm 8-2.
Sample mark line 9; 0.4cm place, sample adsorption fibrage 2 side is partial to by the first diaphragm that the intersection of sample adsorption layer 2 and binding layer 3 is corresponding, and the right-hand member of sample mark line is printed on arrow and MAX printed words,
Wherein, sample adsorption layer 2, binding layer 3, cellulose rete 4, each layer of water accepting layer 5 are pasted onto successively on supporting layer 1, the intersection fiber spliced each other crosses one another infiltration; The spread pattern of detection line and control line is
Embodiment 5
A kind of test strips detecting chicken trachitis virus comprises:
Supporting layer 1, it is made with plastic slice bar;
Detection layers, it comprises the sample adsorption layer 2 connected successively, its fibrage made with glass wool; Binding layer 3, it is the fibrage that the nylon fiber of the anti-ILTV monoclonal antibody being attached with colloid gold label is made; Cellulose rete 4, its cellulose rete made for polyvinylidene fluoride; Water accepting layer 5, it is the water accepting layer be made up of absorbent filter;
Detection line 6; It is on cellulose rete 4, detects trace 6 (code name T) with the monoclonal antibody IgG solution of anti-another epi-position of ILTV mark;
Control line 7; It is on cellulose rete 4, with sheep (rabbit) anti-mouse IgG solution mark contrast trace 7 (code name C);
First diaphragm 8-1; It is the white diaphragm of the test lead covered above sample adsorption fibrage 2 and binding layer 3,
Second diaphragm 8-2; Water accepting layer 5 is coated with powder color diaphragm 8-2.
Sample mark line 9; 0.6cm place, sample adsorption fibrage 2 side is partial to by the first diaphragm that the intersection of sample adsorption layer 2 and binding layer 3 is corresponding, and the right-hand member of sample mark line is printed on arrow and MAX printed words,
Wherein, sample adsorption layer 2, binding layer 3, cellulose rete 4, each layer of water accepting layer 5 are pasted onto successively on supporting layer 1, the intersection fiber spliced each other crosses one another infiltration; The spread pattern of detection line and control line is
Embodiment 6
A kind of test strips detecting chicken trachitis virus comprises:
Supporting layer 1, it is made with the cardboard bar do not absorbed water;
Detection layers, it comprises the sample adsorption layer 2 connected successively, its fibrage made with nylon fiber; Binding layer 3, it is the fibrage that the nylon fiber of the anti-ILTV monoclonal antibody being attached with colloid gold label is made; Cellulose rete 4, its cellulose rete made for pure cellulose film; Water accepting layer 5, it is the water accepting layer be made up of absorbent filter;
Detection line 6; It is on cellulose rete 4, detects trace 6 (code name T) with the monoclonal antibody IgG solution of anti-another epi-position of ILTV mark;
Control line 7; It is on cellulose rete 4, with sheep (rabbit) anti-mouse IgG solution mark contrast trace 7 (code name C);
First diaphragm 8-1; It is the white diaphragm of the test lead covered above sample adsorption fibrage 2 and binding layer 3,
Second diaphragm 8-2; Water accepting layer 5 is coated with powder color diaphragm 8-2.
Sample mark line 9; 0.6cm place, sample adsorption fibrage 2 side is partial to by the first diaphragm that the intersection of sample adsorption layer 2 and binding layer 3 is corresponding, and the right-hand member of sample mark line is printed on arrow and MAX printed words,
Wherein, sample adsorption layer 2, binding layer 3, cellulose rete 4, each layer of water accepting layer 5 are pasted onto successively on supporting layer 1, the intersection fiber spliced each other crosses one another infiltration; The spread pattern of detection line and control line is
Embodiment 7
A kind of test strips detecting chicken trachitis virus comprises:
Supporting layer 1, it is made with the cardboard bar do not absorbed water;
Detection layers, it comprises the sample adsorption layer 2 connected successively, its fibrage made with dacron film; Binding layer 3, it is the fibrage that the nylon fiber of the anti-ILTV monoclonal antibody being attached with colloid gold label is made; Cellulose rete 4, its cellulose rete made for carboxylated cellulose film; Water accepting layer 5, it is the water accepting layer be made up of absorbent filter;
Detection line 6; It is on cellulose rete 4, detects trace 6 (code name T) with the monoclonal antibody IgG solution of anti-another epi-position of ILTV mark;
Control line 7; It is on cellulose rete 4, with sheep (rabbit) anti-mouse IgG solution mark contrast trace 7 (code name C);
First diaphragm 8-1; It is the white diaphragm of the test lead covered above sample adsorption fibrage 2 and binding layer 3,
Second diaphragm 8-2; Water accepting layer 5 is coated with powder color diaphragm 8-2.
Sample mark line 9; 0.6cm place, sample adsorption fibrage 2 side is partial to by the first diaphragm that the intersection of sample adsorption layer 2 and binding layer 3 is corresponding, and the right-hand member of sample mark line is printed on arrow and MAX printed words,
Wherein, sample adsorption layer 2, binding layer 3, cellulose rete 4, each layer of water accepting layer 5 are pasted onto successively on supporting layer 1, the intersection fiber spliced each other crosses one another infiltration; The spread pattern of detection line and control line is
Embodiment 8
A kind of test strips detecting chicken trachitis virus comprises:
Supporting layer 1, it is made with the cardboard bar do not absorbed water;
Detection layers, it comprises the sample adsorption layer 2 connected successively, its fibrage made with nylon fiber; Binding layer 3, it is the fibrage that the nylon fiber of the anti-ILTV monoclonal antibody being attached with colloid gold label is made; Cellulose rete 4, it is the cellulose rete that polyvinylidene fluoride (PVDF) tunica fibrosa is made; Water accepting layer 5, it is the water accepting layer be made up of absorbent filter;
Detection line 6; It is on cellulose rete 4, detects trace 6 (code name T) with the monoclonal antibody IgG solution of anti-another epi-position of ILTV mark;
Control line 7; It is on cellulose rete 4, with sheep (rabbit) anti-mouse IgG solution mark contrast trace 7 (code name C);
First diaphragm 8-1; It is the white diaphragm of the test lead covered above sample adsorption fibrage 2 and binding layer 3,
Second diaphragm 8-2; Water accepting layer 5 is coated with powder color diaphragm 8-2.
Sample mark line 9; 0.6cm place, sample adsorption fibrage 2 side is partial to by the first diaphragm that the intersection of sample adsorption layer 2 and binding layer 3 is corresponding, and the right-hand member of sample mark line is printed on arrow and MAX printed words,
Wherein, sample adsorption layer 2, binding layer 3, cellulose rete 4, each layer of water accepting layer 5 are pasted onto successively on supporting layer 1, the intersection fiber spliced each other crosses one another infiltration; The spread pattern of detection line and control line is " // ".
Embodiment 9
A kind of test strips detecting chicken trachitis virus comprises:
Supporting layer 1, it is made with the cardboard bar do not absorbed water;
Detection layers, it comprises the sample adsorption layer 2 connected successively, its fibrage made with dacron film; Binding layer 3, it is the fibrage that the nylon fiber of the anti-ILTV monoclonal antibody being attached with colloid gold label is made; Cellulose rete 4, its cellulose rete made for pure cellulose film; Water accepting layer 5, it is the water accepting layer be made up of absorbent filter;
Detection line 6; It is on cellulose rete 4, detects trace 6 (code name T) with the monoclonal antibody IgG solution of anti-another epi-position of ILTV mark;
Control line 7; It is on cellulose rete 4, with sheep (rabbit) anti-mouse IgG solution mark contrast trace 7 (code name C);
First diaphragm 8-1; It is the white diaphragm of the test lead covered above sample adsorption fibrage 2 and binding layer 3,
Second diaphragm 8-2; Water accepting layer 5 is coated with color purple diaphragm 8-2.
Sample mark line 9; 0.5cm place, sample adsorption fibrage 2 side is partial to by the first diaphragm that the intersection of sample adsorption layer 2 and binding layer 3 is corresponding, and the right-hand member of sample mark line is printed on arrow and MAX printed words,
Wherein, sample adsorption layer 2, binding layer 3, cellulose rete 4, each layer of water accepting layer 5 are pasted onto successively on supporting layer 1, the intersection fiber spliced each other crosses one another infiltration; The spread pattern of detection line and control line is " // ".
Detection method of operating of the present utility model comprises the following steps:
The preparation of A, detection sample liquid, gets the larynx of disease chicken, tracheal mucosa and transudate etc., is then shredded, grinds, make the detected sample suspension of 1: 2-1: 5 times with physiological saline, puts 4 DEG C of room temperatures clarifications or centrifugal;
B, detection operation, inserted in detected sample by this test strips test lead, insertion depth is no more than sample mark line, and take out test strips after about 30 seconds, horizontal positioned is about 1-5 minute, simultaneously observations.
C, result judge, if only demonstrate brownish red contrast trace C on the cellulose membrane of test strips, represent that surveying inspection result is negative, and illustrate not detect ILTV in test sample liquid; If henna control line C and detection line T appears in the cellulose membrane in test strips, represent that testing result is positive, namely in measuring samples, detect ILTV; If without any the display of brownish red trace on cellulose membrane, then show that test strips had lost efficacy or operated wrong.
The Cleaning Principle of above-mentioned test strips:
After test strips test lead (sample adsorption fibrage) inserts detected sample solution, solution to be checked drives the golden labeling antibody in sick chicken virus to be checked and binding layer to spread to cellulose membrane together by chromatography effect, and finally infiltrate in the water accepting layer of handle end (the second end), the gold mark monoclonal antibody that in diffusion process, virus to be checked can be corresponding with this virus combines, and then the many anti-igg of this virus that resist on cellulose membrane in detection line are combined, thus demonstrate henna detection line T; Goat-anti in control line or rabbit anti-mouse IgG then can mark monoclonal antibody with gold and be combined, and form brownish red control line C.If do not have ILTV in measuring samples liquid, test strips only demonstrates a brownish red control line C; If without any the display of brownish red trace on cellulose membrane, then show that test strips had lost efficacy or misoperation.
The utility model is according to the ultimate principle of ELISA, virus is detected with immune chromatography test paper, the diafiltration of miillpore filter is utilized to concentrate and capillarity, antigen-antibody reaction is forwarded on solid phase filter membrane by the Traditional liquid phase environment of ELISA and carries out fast, and adopt collaurum trace to replace enzyme trace, with the colour developing situation of direct visual perception collaurum, immediately obtain testing result, therefore the method is more easier, quick than serological methods such as ELISA.
The utility model detection specificity is strong, and susceptibility is high.This test strips is prepared from based on the specific monoclonal antibody of collaurum trace high-affinity/many anti-, formed without covalent bond between gold grain and antibody molecule in gold labeling antibody, the two is combined by the Van der Waals force between the charges of different polarity, collaurum mark affects very little on monoclonal antibody/many anti-specificitys and affinity (adhesion), and has higher trace rate.Therefore, this test strips has higher specificity and susceptibility, the albumen of the corresponding virus of 2 nanograms can be detected.And, easy and simple to handle, fast.Without the need to additional any Other Instruments and reagent when using the utility model test strips, as long as its test lead to be inserted in measuring samples liquid about 30 seconds, then testing result can be judged at about 5 minutes.
In addition, use testing result intuitive display of the present utility model, accurately.This test strips is to show henna detection trace and contrast trace as the positive detected and negative trace, and namely on cellulose membrane, show two brownish red traces, represent has virus to detect in detected sample, and result be the positive; Only display brownish red contrast trace C on cellulose membrane, represent and do not detect virus in detected sample liquid, result be feminine gender.Result judges directly perceived, accurate, simple and clear, not easily occurs false negative and false positive erroneous judgement.
Further, the test strips that the utility model provides, can reduce investment and testing cost.Use this test strips, do not need separately to join Other Instruments, equipment and reagent, save large measuring appratus, equipment and additive reagent expense; Specialty and layman all can carry out Site Detection whenever and wherever possible, remove the travelling expenses of diagnosis room without the need to paying expert diagnosis Laboratory Fee or feeding sample, and saving testing cost, testing cost is low.
The utility model is simple to operate, become " single step " or " foolproof ", and be convenient for carrying and preserve, the needs of different levels personnel can be met, comprise specialty chemical examination, customs quarantine control, health and epidemic prevention, quality monitoring, livestock products processing, intensive culture to individual cultivation etc., there are wide market outlook and good economical, societal benefits, applied range, be convenient to popularity application.
Although embodiment of the present utility model is open as above, but it is not restricted to listed in instructions and embodiment utilization, it can be applied to various applicable field of the present utility model completely, for those skilled in the art, can easily realize other amendment, therefore do not deviating under the universal that claim and equivalency range limit, the utility model is not limited to specific details and illustrates here and the legend described.
Claims (10)
1. detect a test strips for chicken trachitis virus, it is characterized in that, comprising:
Detection layers, it comprises the sample adsorption layer, binding layer, cellulose rete and the water accepting layer that connect successively, described sample adsorption layer defines the first end for contacting detected sample, described water accepting layer defines the second end away from detected sample, and described binding layer includes the first antibody of the chicken trachitis virus of colloid gold label;
Detection line, it is arranged on described cellulose rete, and described detection line comprises the second antibody of the chicken trachitis virus of bag quilt, and described first antibody is different with the site that the scorching virus of described second antibody and described chicken trunnion combines;
Control line, it is also arranged on described cellulose rete and than described detection line away from described first end, described control line comprises the sheep/rabbit anti-mouse immunoglobulin G of bag quilt.
2. the test strips detecting chicken trachitis virus as claimed in claim 1, it is characterized in that, described first antibody is monoclonal antibody or the polyclonal antibody of chicken trachitis virus, and described second antibody is polyclonal antibody or the monoclonal antibody of chicken trachitis virus.
3. the test strips of the detection chicken trachitis virus as described in any one of claim 1 to 2, is characterized in that, also comprise:
Supporting layer, described detection layers is fixed on the upper surface of described supporting layer.
4. the test strips detecting chicken trachitis virus as claimed in claim 3, is characterized in that, also comprise:
First diaphragm, it is arranged on above described detection layers, and one end of described first diaphragm bends downwards and envelopes described first end, and the other end extends to the top of described cellulose rete one end;
Second diaphragm, it is arranged on the top of described detection layers, and one end of described second diaphragm bends downwards and envelopes described second end, and the other end extends to the top of the described cellulose rete other end;
Wherein, described first diaphragm is white, and the color of described second diaphragm is not identical with the color of described first diaphragm.
5. the test strips detecting chicken trachitis virus as claimed in claim 4; it is characterized in that; described first diaphragm is provided with sample mark line, and described sample mark line is 0.3 ~ 0.7cm place with the distance away from the side of the described sample adsorption layer of described first end.
6. the as claimed in claim 1 test strips detecting chicken trachitis virus, is characterized in that, described cellulose rete is nitrocellulose filter, any one in cellulose membrane, carboxylated cellulose film and PVDF membrane make.
7. the test strips detecting chicken trachitis virus as claimed in claim 1, it is characterized in that, described sample adsorption layer is made up of any one in glass wool, nylon fiber and dacron.
8. the test strips detecting chicken trachitis virus as claimed in claim 1, it is characterized in that, described binding layer is made up of any one in glass wool, nylon fiber and dacron.
9. the test strips detecting chicken trachitis virus as claimed in claim 4, it is characterized in that, described supporting layer is made up of the hard plastic sheet not absorbing water or cardboard bar; Described water accepting layer thieving paper is made.
10. the as claimed in claim 1 test strips detecting chicken trachitis virus, is characterized in that, the spread pattern of described detection line and described control line be " || ", " ++ ", "
", "
" and "
" in any one.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201520097970.8U CN204495834U (en) | 2015-02-11 | 2015-02-11 | Detect the test strips of chicken trachitis virus |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201520097970.8U CN204495834U (en) | 2015-02-11 | 2015-02-11 | Detect the test strips of chicken trachitis virus |
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| Publication Number | Publication Date |
|---|---|
| CN204495834U true CN204495834U (en) | 2015-07-22 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201520097970.8U Expired - Fee Related CN204495834U (en) | 2015-02-11 | 2015-02-11 | Detect the test strips of chicken trachitis virus |
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| CN (1) | CN204495834U (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105891489A (en) * | 2016-04-08 | 2016-08-24 | 王秀梅 | Test paper for detecting HBcAb (hepatitis B core antibody) IgM (immunoglobulin m) in serum and preparation method of test paper |
| CN110146702A (en) * | 2019-06-19 | 2019-08-20 | 四川省畜牧科学研究院 | A kind of test strips detecting infectious laryngotracheitis virus, preparation method, detection method and kit |
-
2015
- 2015-02-11 CN CN201520097970.8U patent/CN204495834U/en not_active Expired - Fee Related
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105891489A (en) * | 2016-04-08 | 2016-08-24 | 王秀梅 | Test paper for detecting HBcAb (hepatitis B core antibody) IgM (immunoglobulin m) in serum and preparation method of test paper |
| CN110146702A (en) * | 2019-06-19 | 2019-08-20 | 四川省畜牧科学研究院 | A kind of test strips detecting infectious laryngotracheitis virus, preparation method, detection method and kit |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20150722 Termination date: 20160211 |
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| CF01 | Termination of patent right due to non-payment of annual fee |