CN111537734A - Duck variant parvovirus N protein IgY antibody colloidal gold detection test paper and preparation method and application thereof - Google Patents

Duck variant parvovirus N protein IgY antibody colloidal gold detection test paper and preparation method and application thereof Download PDF

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CN111537734A
CN111537734A CN202010409998.6A CN202010409998A CN111537734A CN 111537734 A CN111537734 A CN 111537734A CN 202010409998 A CN202010409998 A CN 202010409998A CN 111537734 A CN111537734 A CN 111537734A
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卞传忠
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Anhui Zhongqi Biotechnology Co ltd
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Abstract

The invention relates to a duck variant parvovirus N protein IgY antibody colloidal gold test paper and a preparation method and application thereof, belonging to the technical field of rapid detection of duck variant parvovirus N protein IgY antibodies. The test strip can rapidly detect the level of the duck variant parvovirus N protein IgY antibody, and compared with a conventional ELISA antibody detection method, the test strip is more convenient and rapid.

Description

Duck variant parvovirus N protein IgY antibody colloidal gold detection test paper and preparation method and application thereof
Technical Field
The invention belongs to the technical field of rapid detection of duck variant parvovirus N protein IgY antibodies, and particularly relates to a duck variant parvovirus N protein IgY antibody colloidal gold detection test paper and a preparation method and application thereof.
Background
Since 2014, many meat duck farms in the east of China have developed a new duck viral infectious disease, which is clinically mainly characterized by slow growth and development of ducklings, atrophy of upper and lower beaks, swelling and extending of duck tongues and difficulty in walking. The temporary name is duck short beak dwarfism syndrome, which is caused by duck variant parvovirus. The fatality rate of the disease is very low, but the morbidity rate is 10% -30%, and the morbidity rate of part of duck groups reaches 50%. Causing serious economic loss to the duck breeding industry. The duck infectious disease is a newly appeared duck infectious disease in China.
The colloidal gold immunochromatography technology is a mature and widely applied immune labeling technology after the traditional three immune labeling technologies. When a sample to be detected is added on a sample membrane, the antigen-antibody reaction is rapidly carried out, the detection generally takes 5-15 min to obtain a result, and compared with other methods (such as ELISA, 1-2 h is needed), the detection time is greatly shortened. The test result is judged by a macroscopic color development strip, special instruments and equipment are not needed, only a test strip or a percolation kit is needed, and the sample is only subjected to very simple treatment or no pretreatment. The method has the advantages of low cost, simple operation, reagent stability, no influence of external factors such as temperature and the like, convenience, rapidness, specificity, sensitivity, strong stability, intuitive result judgment and the like, is particularly suitable for on-site rapid inspection, has huge development potential and wide application prospect, and represents the development direction of simple, rapid and convenient popularization of diagnostic reagents.
At present, the detection method aiming at duck variant parvovirus antibody is mainly enzyme-linked immunosorbent assay, and colloidal gold is also related in the field of parvovirus antibody detection, such as: chinese patent document CN201620169992.5 discloses a test strip for rapidly detecting Muscovy duck parvovirus antibodies, which comprises a supporting layer, an adsorption fiber layer, a water absorption layer, a cellulose membrane layer and a gold-labeled fiber layer, wherein the cellulose membrane layer is provided with a detection line and a quality control line, the detection line is marked with purified Muscovy duck parvovirus antibodies, the quality control line is marked with sheep or rabbit anti-Muscovy duck parvovirus antibodies, and the gold-labeled fiber layer is attached with Muscovy duck parvovirus marked by colloidal gold or recombinant Muscovy duck parvovirus VP2 protein marked by colloidal gold; chinese patent document CN201610883318.8 discloses a colloidal gold test paper capable of simultaneously detecting waterfowl-derived parvovirus and a preparation method thereof, wherein a gold-labeled pad is coated with a colloidal gold-labeled protein with a gosling plague virus resistant monoclonal antibody I, a detection line is coated with a gosling plague virus resistant monoclonal antibody II, and a quality control line is coated with a goat anti-mouse IgG antibody. However, in both of the above documents, antibody-coated colloidal gold is adopted, and the invention prepares a colloidal gold test strip for detecting the duck variant parvovirus N protein IgY antibody according to a competitive inhibition method, and compared with the colloidal gold coated with the antibody, the coated antigen detection is more accurate, faster and more stable.
Disclosure of Invention
The invention aims to solve the problems and provide a duck variant parvovirus N protein IgY antibody, a preparation method and an application thereof, so as to provide a more accurate, rapid and stable rapid detection method for the duck variant parvovirus N protein IgY antibody.
The invention realizes the purpose through the following technical scheme:
the invention provides a duck variant parvovirus N protein IgY antibody colloidal gold test paper which comprises a PVC (polyvinyl chloride) bottom plate, a sample pad, a combination pad, a nitrocellulose membrane and absorbent filter paper, wherein the combination pad is a colloidal gold pad coated with a duck variant parvovirus N protein-colloidal gold compound, the nitrocellulose membrane is provided with a quality control line (C line) and a detection line (T line), the quality control line is coated with a rabbit anti-chicken IgY antibody, and the detection line is coated with a mouse anti-duck variant parvovirus N protein IgG antibody.
The invention also provides a preparation method of the test paper, which comprises the following steps:
(1) preparing purified duck variant parvovirus N protein;
(2) preparing a mouse anti-duck variant parvovirus N protein IgG antibody;
(3) respectively diluting rabbit anti-chicken IgY antibody (purchased from Solebao science and technology Co., Ltd.: SPA235) and mouse anti-duck variant parvovirus N protein IgG antibody, and coating on a quality control line and a detection line of a nitrocellulose membrane;
(4) preparing a duck variant parvovirus N protein-colloidal gold compound, and coating the compound on a bonding pad to obtain a colloidal gold pad;
(5) and (6) assembling.
As a further optimization scheme of the invention, the step (1) of preparing the purified duck variant parvovirus N protein comprises the following steps: preparing an escherichia coli recombinant strain of duck variant parvovirus N protein according to a known duck variant parvovirus N protein gene sequence, fermenting and expressing the recombinant strain, centrifuging to obtain a strain, crushing the strain, centrifuging to obtain a supernatant, and passing the supernatant through an affinity chromatography column to obtain the purified duck variant parvovirus N protein.
As a further optimization scheme of the invention, the step (2) of preparing the mouse anti-duck variant parvovirus N protein IgG antibody comprises the following steps: mixing and emulsifying the purified duck variant parvovirus N protein and an immunologic adjuvant completely, then carrying out intraperitoneal injection on the mice, enhancing immunity every 2-3 weeks, cutting off the heads and taking blood after the antibody titer reaches a set level, and centrifuging to obtain the serum containing the mouse anti-duck variant parvovirus N protein IgG antibody.
As a further optimization scheme of the invention, in the step (3), the rabbit anti-chicken IgY antibody and the mouse anti-duck variant parvovirus N protein IgG antibody are respectively diluted by 50mM PBS to the final concentration of 1mg/ml, then sprayed on the quality control line and the detection line in the spraying amount of 1 muL/cm, and fully dried to finish coating;
as a further optimized scheme of the present invention, in the step (4), the preparation method of the duck variant parvovirus N protein-colloidal gold complex comprises the following steps: dissolving the purified duck variant parvovirus N protein in 0.2-0.4ml of purified water, and adjusting the pH value of the colloidal gold solution to 7.0; mixing duck variant parvovirus N protein with colloidal gold according to the following combination: adding 10ml of colloidal gold solution into every 10mg of duck variant parvovirus N protein; mixing for 30min, adding BSA blocking solution with final concentration of 1%, reacting at room temperature for 30min, centrifuging at low speed of 2000r/min for 10min, and centrifuging at high speed of 12000r/min for 30 min; using a protective agent containing: 0.1% sucrose and 0.1% trehalose, activating agent: and (3) carrying out heavy suspension precipitation on the Tris-HCL (pH7.0) solution of 0.2 percent of PEG6000 to obtain the duck variant parvovirus N protein-colloidal gold compound.
As a further optimization scheme of the invention, in the step (4), the duck variant parvovirus N protein-colloidal gold complex is coated on the bonding pad in a spraying amount of 1 mu L/cm to obtain the colloidal gold pad.
As a further optimization scheme of the invention, the assembling step of the step (5) comprises the following steps: preparing a sample pad, a colloidal gold pad, a coated nitrocellulose membrane, absorbent filter paper and a PVC bottom plate in a dry environment, sticking the coated nitrocellulose membrane at the center of the PVC bottom plate, sticking the absorbent filter paper on the upper edge of the nitrocellulose membrane, sticking the colloidal gold pad on the lower edge of the nitrocellulose membrane, sticking the sample pad on the lower edge of the colloidal gold pad, and cutting the test paper plate into strips after the completion.
The invention also provides a method for detecting the duck variant parvovirus N protein IgY antibody by using the test paper, which comprises the following steps: the test paper is horizontally placed, 50-10 mu L of a sample to be tested is taken, a sample pad is added, and after standing for 15min at room temperature, the result is judged according to the following conditions:
the test paper is effective: the test paper is proved to be effective when a purple line appears on a quality control line (line C);
negative: a purple red line appears on the quality control line (C line) and the detection line (T line);
positive: only a purple red line appears on a quality control line (C line), and a color line does not appear on a detection line (T line);
and the shade of the color line is in direct proportion to the antibody titer, i.e. the higher the antibody titer, the lighter the color line of the detection line (T line).
The invention has the beneficial effects that: the invention provides a duck variant parvovirus N protein IgY antibody colloidal gold test paper and a preparation method and application thereof, after the test paper is used for detection, a positive result shows that a detected sample duck variant parvovirus N protein IgY antibody is positive and reaches a required antibody level, a negative result shows that the sample duck variant parvovirus N protein IgY antibody is negative or does not reach the required antibody level, and the test paper can rapidly detect the duck variant parvovirus N protein IgY antibody level.
Drawings
FIG. 1 is a schematic structural diagram of a duck variant parvovirus N protein IgY antibody immune colloidal gold test strip.
Detailed Description
The present application will now be described in further detail with reference to the drawings, it should be noted that the following detailed description is given for illustrative purposes only and is not to be construed as limiting the scope of the present application, as those skilled in the art will be able to make numerous insubstantial modifications and adaptations to the present application based on the above disclosure.
Example 1
The preparation method of the duck variant parvovirus N protein IgY antibody colloidal gold test paper provided by the embodiment comprises the following steps:
(1) preparation of purified duck variant parvovirus N protein
Preparing an escherichia coli recombinant strain of duck variant parvovirus N protein according to a known gene sequence (GENBANK accession number: NC-006147.2) of the duck variant parvovirus N protein, fermenting and expressing the recombinant strain, centrifuging to obtain a strain, crushing the strain, centrifuging to obtain a supernatant, and passing the supernatant through an affinity chromatography column to obtain the purified duck variant parvovirus N protein.
(2) Preparation of mouse anti-duck variant parvovirus N protein IgG antibody
Mixing and emulsifying the purified duck variant parvovirus N protein and an immunologic adjuvant completely, then carrying out intraperitoneal injection on the mice, enhancing the immunity every 2-3 weeks, taking blood after the heads break after the antibody titer reaches an ideal level, and centrifuging to obtain the mouse anti-duck variant parvovirus N protein IgG antibody serum.
(3) Preparation of NC Membrane 4 (nitrocellulose Membrane)
The rabbit anti-chicken IgY antibody (purchased from Solebao science and technology Co., Ltd.: SPA235) and the mouse anti-duck variant parvovirus N protein IgG antibody are respectively diluted with 50mM PBS to a final concentration of 1mg/ml, and sprayed on a quality control line (C line) and a detection line (T line) with a spraying amount of 1 muL/cm, and fully dried.
(4) Preparation of the colloidal gold pad 3
Preparing 20-50nm colloidal gold solution by a trisodium citrate reduction method: taking HAuCl with the mass fraction of 0.01-0.02%4Heating 100ml water solution, boiling, rapidly adding 1ml trisodium citrate water solution with mass fraction of 1% -5%, boiling for about 5min to obtain gold particle solution with particle diameter of 20-50nmThe yellowish aqueous chloroauric acid solution was observed to turn gray, then black, and then gradually stabilized to wine-red color after the addition of trisodium citrate, and then cooled to room temperature and stored at 2-8 ℃ in the dark.
Dissolving the purified duck variant parvovirus N protein in 0.2-0.4ml of purified water, and adjusting the pH value of the colloidal gold solution to 7.0; mixing duck variant parvovirus N protein with colloidal gold according to the following combination: adding 10ml of colloidal gold solution into every 10mg of duck variant parvovirus N protein; mixing for 30min, adding blocking solution (BSA with final concentration of 1%) at room temperature, allowing to act and bind for 30min, centrifuging at low speed of 2000r/min for 10min, and centrifuging at high speed of 12000r/min for 30 min; using a protective agent containing: 0.1% sucrose and 0.1% trehalose, activating agent: and (3) carrying out heavy suspension precipitation on the Tris-HCL (pH7.0) solution of 0.2 percent of PEG6000 to obtain the duck variant parvovirus N protein-colloidal gold compound.
And (3) uniformly spraying the prepared duck variant parvovirus N protein-colloidal gold compound on a glass cellulose membrane according to the spraying amount of 1 mu L/cm, and fully drying to obtain the colloidal gold pad 3.
(5) Assembly of test strips
Preparing water-absorbing filter paper 5, a sample pad 2 and a PVC base plate 1 in a drying room, as shown in figure 1, pasting the NC membrane 4 coated in the step (2) on the center of the PVC base plate 1, pasting the water-absorbing filter paper 5 on the lower edge of the chromatography direction d ' of the NC membrane 4, pasting the colloidal gold pad 3 prepared in the step (4) on the upper edge of the chromatography direction d ' of the NC membrane 4, pasting the sample pad 2 on the upper edge of the chromatography direction d ' of the colloidal gold pad 3, wherein the overlapped parts of the components are 1-2mm, and cutting the pasted test paper plate into test paper strips with the width of 3mm by using a cutting machine after the completion. And (3) putting the cut test strip into a card shell, and sealing the test strip and the drying agent in an aluminum foil bag to complete the assembly of the product.
According to the antibody level required by the delivery of the duck variant parvovirus N protein IgY antibody, taking the duck variant parvovirus N protein IgY antibody, diluting the duck variant parvovirus N protein IgY antibody to 50ng/ml concentration, 100ng/ml concentration, 200ng/ml concentration, 400ng/ml concentration and 800ng/ml concentration by using 50mM PBS solution as a detection sample, setting a blank control, and detecting the antibody concentration by respectively using the test strip and an enzyme-linked immunosorbent assay (ELISA) of the invention, wherein the detection sample specifically comprises the following steps: the test strip is flatly placed on a desktop, 50-100 mu L of serum sample to be tested is taken, a sample pad is added, and after standing for 15min at room temperature, the result is judged according to the following conditions:
the test paper is effective: the test paper is proved to be effective when a purple line appears on a quality control line (line C);
negative: a purple red line appears on the quality control line (C line) and the detection line (T line);
positive: only a purple red line appears on a quality control line (C line), and a color line does not appear on a detection line (T line);
and the shade of the color line is in direct proportion to the antibody titer, i.e. the higher the antibody titer, the lighter the color line of the detection line (T line).
The concentration of the antibody is detected by enzyme-linked immunosorbent assay (ELISA) according to the conventional ELISA method.
The results are shown in table 1 below:
table 1: method for detecting concentration of duck variant parvovirus N protein IgY antibody by different methods
Figure BDA0002492810950000051
Table 1 shows that the detection result of the colloidal gold test strip of the present invention is highly consistent with the ELISA detection result, which indicates that the accuracy of the colloidal gold test strip of the present invention is 100%, and it is feasible to detect the duck variant parvovirus N protein IgY antibody and the content thereof using the colloidal gold test strip of the present invention. Because the depth of the color line is inversely proportional to the antibody titer, the higher the antibody titer is, the lighter the color line is, whether the commercial duck variant parvovirus N protein IgY antibody product reaches the delivery antibody level can be detected, so as to judge whether the purchased product is a fake product or adulterated.
The invention uses the coated antigen colloidal gold, and the detection is more accurate compared with the colloidal gold coated with the antibody, and the specific content is as follows.
Coated antibody colloidal gold example:
(1) preparation of purified duck variant parvovirus N protein
Consistent with the competitive inhibition method
(2) Preparation of NC Membrane 4 (nitrocellulose Membrane)
Goat anti-rabbit IgG (purchased from Solebao scientific and technology Co., Ltd.: SPA134) and purified duck variant parvovirus N protein were respectively diluted with 50mM PBS to a final concentration of 1mg/ml, and then uniformly sprayed on a quality control line (C line) and a detection line (T line) on an NC membrane (nitrocellulose membrane) at a spraying amount of 1. mu.L/cm, followed by sufficient drying.
(3) Preparation of the colloidal gold pad 3
The preparation method of the 20-50nm colloidal gold by using a trisodium citrate reduction method comprises the following specific operation methods: taking 100ml of HAuCl4 aqueous solution with the mass fraction of 0.01-0.02%, heating and boiling, quickly adding 1ml of trisodium citrate aqueous solution with the mass fraction of 1-5%, continuously boiling for about 5min to prepare gold particles with the particle size of 20-50nm, wherein the yellowish chloroauric acid aqueous solution can be observed to quickly turn into gray after the trisodium citrate is added, then turn into black, gradually stabilize into wine red, and be stored away from light at the temperature of 2-8 ℃ after being cooled to room temperature.
Dissolving high-purity rabbit anti-chicken IgY (purchased from Solebao science and technology Co., Ltd.: SPA235) in 0.2-0.4ml of purified water, and adjusting the pH value of the colloidal gold solution to 7.0; rabbit anti-chicken IgY was mixed with colloidal gold according to the following combinations: adding 10ml of colloidal gold solution into every 10mg of rabbit anti-chicken IgY; mixing for 30min, adding blocking solution (BSA with final concentration of 1%) at room temperature, allowing to act and bind for 30min, centrifuging at low speed of 2000r/min for 10min, and centrifuging at high speed of 12000r/min for 30 min; using a protective agent containing: 0.1% sucrose and 0.1% trehalose, activating agent: and (3) carrying out heavy suspension precipitation on the solution of 0.2 percent PEG6000 in Tris-HCL (pH7.0), thus obtaining the rabbit anti-chicken IgY-colloidal gold compound.
The rabbit anti-chicken IgY-colloidal gold compound prepared above is uniformly sprayed on a glass cellulose membrane according to the spraying amount of 1 muL/cm, and is fully dried to obtain the binding pad 3.
(4) Assembly of test strips
Preparing water-absorbing filter paper 5, a sample pad 2 and a PVC base plate 1 in a drying room, as shown in figure 1, pasting the NC membrane 4 coated in the step (2) on the center of the PVC base plate 1, pasting the water-absorbing filter paper 5 on the lower edge of the chromatography direction d ' of the NC membrane 4, pasting the colloidal gold pad 3 prepared in the step (4) on the upper edge of the chromatography direction d ' of the NC membrane 4, pasting the sample pad 2 on the upper edge of the chromatography direction d ' of the colloidal gold pad 3, wherein the overlapped parts of the components are 1-2mm, and cutting the pasted test paper plate into test paper strips with the width of 3mm by using a cutting machine after the completion. And (3) putting the cut test strip into a card shell, and sealing the test strip and the drying agent in an aluminum foil bag to complete the assembly of the product.
The provided detection test paper using and result judging method can be carried out according to the following steps: the test strip is flatly placed on a desktop, 50-100 mu L of serum sample to be tested is taken, a sample pad is added, and after standing for 15min at room temperature, the result is judged according to the following conditions:
the test paper is effective: the test paper is proved to be effective when a purple line appears on a quality control line (line C);
negative: only a purple red line appears on a quality control line (C line), and a color line does not appear on a detection line (T line);
positive: a purple red line appears on the quality control line (C line) and the detection line (T line);
and the shade of the color line is directly proportional to the antibody titer, i.e., the higher the antibody titer, the deeper the color line.
Test strip detection effect comparison of two coating methods
Table 2: method for detecting concentration of duck variant parvovirus N protein IgY antibody by different methods
Figure BDA0002492810950000071
Figure BDA0002492810950000081
As can be seen from Table 2, the test results of the coated antigen colloidal gold test strip of the present invention are more accurate than the test results of the coated antibody colloidal gold test strip.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the present invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention.

Claims (9)

1. The colloidal gold test paper for the duck variant parvovirus N protein IgY antibody comprises a PVC bottom plate, a sample pad, a combination pad, a nitrocellulose membrane and absorbent filter paper, wherein the nitrocellulose membrane is provided with a quality control line and a test line.
2. A method of making the test strip of claim 1, comprising the steps of:
(1) preparing purified duck variant parvovirus N protein;
(2) preparing a mouse anti-duck variant parvovirus N protein IgG antibody;
(3) respectively diluting rabbit anti-chicken IgY antibody and mouse anti-duck variant parvovirus N protein IgG antibody, and coating the diluted antibodies on a quality control line and a detection line of a nitrocellulose membrane;
(4) preparing a duck variant parvovirus N protein-colloidal gold compound, and coating the compound on a bonding pad to obtain a colloidal gold pad;
(5) and (6) assembling.
3. The method for preparing test paper according to claim 2, wherein the step (1) of preparing purified duck variant parvovirus N protein comprises: preparing an escherichia coli recombinant strain of duck variant parvovirus N protein according to a known duck variant parvovirus N protein gene sequence, fermenting and expressing the recombinant strain, centrifuging to obtain a strain, crushing the strain, centrifuging to obtain a supernatant, and passing the supernatant through an affinity chromatography column to obtain the purified duck variant parvovirus N protein.
4. The method for preparing test paper according to claim 2, wherein the step (2) of preparing the mouse anti-duck variant parvovirus N protein IgG antibody comprises the steps of: mixing and emulsifying the purified duck variant parvovirus N protein and an immunologic adjuvant completely, then carrying out intraperitoneal injection on the mice, enhancing immunity every 2-3 weeks, cutting off the heads and taking blood after the antibody titer reaches a set level, and centrifuging to obtain the serum containing the mouse anti-duck variant parvovirus N protein IgG antibody.
5. The method for preparing test paper according to claim 2, wherein in the step (3), the rabbit anti-chicken IgY antibody and the mouse anti-duck variant parvovirus N protein IgG antibody are respectively diluted with 50mM PBS to a final concentration of 1mg/ml, and then sprayed on the quality control line and the detection line at a spraying amount of 1 μ L/cm, and are fully dried to complete the coating.
6. The method for preparing test paper according to claim 2, wherein in the step (4), the method for preparing the duck variant parvovirus N protein-colloidal gold complex comprises the following steps: dissolving the purified duck variant parvovirus N protein in 0.2-0.4ml of purified water, and adjusting the pH value of the colloidal gold solution to 7.0; mixing duck variant parvovirus N protein with colloidal gold according to the following combination: adding 10ml of colloidal gold solution into every 10mg of duck variant parvovirus N protein; mixing for 30min, adding BSA blocking solution with final concentration of 1%, reacting at room temperature for 30min, centrifuging at low speed of 2000r/min for 10min, and centrifuging at high speed of 12000r/min for 30 min; using a protective agent containing: 0.1% sucrose and 0.1% trehalose, activating agent: and (3) carrying out heavy suspension precipitation on the Tris-HCL (pH7.0) solution of 0.2 percent of PEG6000 to obtain the duck variant parvovirus N protein-colloidal gold compound.
7. The method for preparing test paper according to claim 2, wherein in the step (4), the duck variant parvovirus N protein-colloidal gold complex is coated on the conjugate pad in a spraying amount of 1 μ L/cm to obtain the colloidal gold pad.
8. The method for preparing a test strip according to claim 2, wherein the assembling step of step (5) comprises: preparing a sample pad, a colloidal gold pad, a coated nitrocellulose membrane, absorbent filter paper and a PVC bottom plate in a dry environment, sticking the coated nitrocellulose membrane at the center of the PVC bottom plate, sticking the absorbent filter paper on the upper edge of the nitrocellulose membrane, sticking the colloidal gold pad on the lower edge of the nitrocellulose membrane, sticking the sample pad on the lower edge of the colloidal gold pad, and cutting the test paper plate into strips after the completion.
9. A method for detecting duck variant parvovirus N protein IgY antibody by using the test paper of claim 1, which is characterized by comprising the following steps: the test paper is horizontally placed, 50-10 mu L of a sample to be tested is taken, a sample pad is added, and after standing for 15min at room temperature, the result is judged according to the following conditions:
the test paper is effective: the test paper is proved to be effective when a purple line appears on the quality control line;
negative: a purple red line appears on both the quality control line and the detection line;
positive: only a purple red line appears on a quality control line, and a color line does not appear on a detection line;
and the color line depth is in direct proportion to the antibody titer, namely the higher the antibody titer is, the lighter the color line of the detection line is.
CN202010409998.6A 2020-05-15 2020-05-15 Duck variant parvovirus N protein IgY antibody colloidal gold detection test paper and preparation method and application thereof Withdrawn CN111537734A (en)

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