Dot-immunogold filtration reagent kit for livestock schistosomiasis diagnosis, its preparation and application method

A spot gold diafiltration and schistosomiasis technology, which is applied in the field of livestock schistosomiasis spot gold immune diafiltration diagnostic kit and its preparation, can solve the problems of long processing period, unfavorable industrial development, high detection cost, etc., and achieve stability and reproducibility are good, conducive to industrial development, the effect of simple production process

Inactive Publication Date: 2005-11-23
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The main problem of this method is that it is necessary to prepare schistosomiasis recombinant protein polyclonal antibody or rabbit anti-human IgG or rabbit anti-schistosome egg polyclonal antibody or GST-PcAb. The preparation of these specific polyclonal antibodies has a long cycle, complicated procedures, and high cost, which limits They are popularized and applied in the census of livestock schistosomiasis
Moreover, when preparing colloidal gold-labeled probes, the amount of polyclonal antibody is usually determined by visual inspection of the test tube. In order to ensure sufficient coating and avoid the interference of excess colloidal gold and non-specific protein binding to the detection result judgment, it is generally added on this basis. 10% to 20% of the polyclonal antibody dosage, and then in order to remove the excess free polyclonal antibody, purification such as chromatography or high-speed centrifugation is required. These treatments either require high equipment conditions, or the processing cycle is long, and the steps Complicated, so the cost is high, which is not conducive to industrial development; third, because the method uses the capture polyclonal antibody as the solid phase, and the serum to be tested and the colloidal gold probe as the mobile phase, one reaction box can only detect blood samples from one person , the detection cost is high and the efficiency is low; at the same time, the test box containing the nitrocellulose membrane coated with antigen or polyclonal antibody needs to be stored at 4-8°C, which increases the difficulty for large-scale application; the fourth is There are many reaction steps, and the interference is relatively increased

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: (Diagnostic Kit)

[0029] Including box body, built-in reaction box, black reagent bottle, normal saline bottle (containing 0.85% normal saline), glass capillary tube (inner diameter 1mm) for spotting blood samples and standard positive blood paper and negative blood paper. The purpose of these two papers is provided. One is for the user to check whether the colloidal gold used for schistosomiasis antigen is effective, and the other is to provide the user with a reference standard for distinguishing negative and positive reactions; among them, the reaction box is a small plastic box of 3cm×2.5cm×0.6cm, divided into boxes The bottom and the lid of the box have two parts. There is a 0.8cm small round hole in the center of the lid. The box is filled with absorbent material. Place a layer of 1.1×1.1cm nitrocellulose membrane (aperture diameter) under the small round hole and on top of the absorbent material. 0.45μ or 0.65μ), close the lid tightly to form a reaction b...

Embodiment 2

[0030] Example 2: (Preparation method of colloidal gold 1)

[0031] The present invention uses 1% chloroauric acid, 1% trisodium citrate, 1% tannic acid, 25mMK 2 CO 3 With double distilled water as raw material, colloidal gold is prepared by tannic acid-sodium citrate reduction method. Sodium citrate is mainly a reducing agent, while tannic acid has the dual functions of reduction and protection, controlling the formation process of "crystal nucleus". The amount of tannic acid determines the size of the colloidal gold particle size, so changing the amount of tannic acid can achieve the purpose of preparing colloidal gold particles with the required diameter. In this example, the ratio of each raw material component of liquid A and liquid B is shown in Table 1:

[0032]

[0033] Heat the prepared liquid A and B in a water bath to 59~61℃ at the same time, stir A liquid on an electromagnetic stirrer, quickly add liquid B, continue to stir for 1 minute, and heat to boiling w...

Embodiment 3

[0034] Example 3: (Preparation method of colloidal gold 2)

[0035]

[0036] The preparation and synthesis methods of liquid A and liquid B are the same as in Example 1, and the diameter of the prepared colloidal gold particles is 10

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Abstract

The invention provides a preparing and application method of diffusion medium box for diagnosing livestock's snail fever spot gold, which is a new immune labeling technique and livestock's snail fever immunology diagnosis technique. The invention labels colloid gold onto Japanese snail antigen protein, and is suit for testing many kinds of animals such as cattle, sheep, pig and rabbit; it spots tested animal's blood dried paper leachate onto pyroxylin film as posited phase, then it droplets snail antibody colloid gold 2í½3 drops (100í½150ª–l) and produces naked eye visible red speckle in 2 minutes. Its positive coincidence rate is 100% and its negative coincidence rate is 99.2%.

Description

Technical field [0001] The present invention relates to the technical field of diagnosis of livestock schistosomiasis, in particular to a dot immunofiltration diagnostic kit for livestock schistosomiasis, and a preparation method and application method thereof. Background technique [0002] Domestic animal schistosomiasis not only harms the health of domestic animals, but also is an important spreader of its disease source. With the in-depth development of schistosomiasis control in our country, although great achievements have been made, there are still more than 100 counties (cities, districts) in our country that have failed to control the spread of the disease. At the same time, the disease has basically been eliminated in provinces and regions. There is still the task of long-term consolidation and monitoring. If the conventional fecal-hatched Schistosomiasis trichome method is used for detection, a large investment of manpower, material resources and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569
Inventor 张雪娟卢福庄方兰勇冯尚连项美华程菊芬周永学
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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