Method for preparing porcine circovirus type 2 colloidal gold antibody fast test strip

A technology for porcine circovirus detection and test paper, which is applied in the field of veterinary biology and can solve the problems of low specificity, long time required for detection, and complicated operation process.

Inactive Publication Date: 2010-11-10
QINGDAO AGRI UNIV +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the antigen preparation process of this method must go through cumbersome modes such as virus cell culture and virus purification. The specificity is not high, the stability is poor, and the cost is high; the detection requ

Method used

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  • Method for preparing porcine circovirus type 2 colloidal gold antibody fast test strip
  • Method for preparing porcine circovirus type 2 colloidal gold antibody fast test strip
  • Method for preparing porcine circovirus type 2 colloidal gold antibody fast test strip

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Experimental program
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Effect test

preparation example Construction

[0074] 2.5 Preparation of expressed protein anti-mouse serum and IFA results

[0075] After immunizing the mice with the expressed protein for 3 times, the blood was collected from the sinus under the frame, and the serum was analyzed. Diluted with PBS 1:20, and infected PK15 cells with PCV2JXL for IFA, the cytoplasm and membrane-specific fluorescence could be seen in the dark field, in the form of hollow circles, and occasionally appeared in the nucleus. IFA was done with virus-inoculated cells and 1:20 diluted normal BALB / c mouse serum, or PBS was used instead of the primary antibody to interact with normal cells, no fluorescence appeared, indicating that the anti-serum contained anti-virus specific antibodies. See Image 6

[0076] 2.6 ELISA reactivity of recombinant expressed protein with PCV2 infected pig serum

[0077] pGEX-PCV 421-37 and pPRO-Rep recombinant bacterial protein after ultrasonic disruption, coated with microtiter plates, and positive serum and negative...

Embodiment

[0082] The porcine circovirus type 2 colloidal gold antibody rapid detection test strip of the present invention has a structure consisting of a water-absorbing pad 1, a nitrocellulose membrane 2, a glass fiber membrane 3 containing a colloidal gold-labeled antigen, a gold-labeled antigen protective film 4, and a reaction The support is composed of a PVC plate 5, wherein the nitrocellulose membrane 2 is set in the middle of the reaction support PVC plate 5, and the nitrocellulose membrane 2 is provided with two color-developing areas T and C, wherein the T area is coated with genetically engineered pigs. Pseudorabies expresses gE protein; the C area is coated with rabbit anti-swine pseudorabies IgG, and on the upper surface of the left end of the nitrocellulose membrane 2T area, a glass fiber membrane 3 containing colloidal gold-labeled antigen is arranged, and the glass fiber membrane 3 containing colloidal gold-labeled antigen The upper surface of the fiber membrane 3 is prov...

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Abstract

The invention relates to a method for preparing a porcine circovirus type 2 colloidal gold antibody fast test strip. In the method, a porcine circovirus type 2 ORF2 protein is expressed by utilizing genetic engineering, and the test strip is prepared by the principle of enzyme-linked immunoassay and membrane chromatography to fast test antibodies in the blood or serum of pigs. The test strip can be widely used for clinically testing porcine circovirus diseases, has no cross reaction with other viruses, and obtains test results 98.63 percent and 95.83 percent consistent with those obtained by the two methods of the ELISA and neutralization test. Compared with the ELISA, a recombinant antigen immune colloidal gold has the remarkable advantages of high security, no need of culturing the viruses per se, the avoidance of virus spread caused by the operation of the viruses, large-batch preparation, simple process, low production cost, stable and homogeneous antigen components, simple, convenient and labor-saving operation, no apparatus, high detection result specificity, high repeatability, the short time of 15 minutes for the whole test, simple, convenient, fast and accurate operation, high sensitivity, intuition and easy result judgment.

Description

technical field [0001] The invention relates to a diagnostic technique in the field of veterinary biotechnology, in particular to a preparation method for a rapid detection test strip of porcine circovirus type 2 colloidal gold antibody. Background technique [0002] Porcine circovirus 2 (PCV2) is the causative agent of post-weaning multisystemic wasting syndrome (PMWS) in weaned pigs, and has been widely recognized. With the in-depth research on PCV2 and its pathogenicity, it has recently been found that PCV2 is also associated with reproductive disorders in pigs, respiratory diseases in weaned pigs and fattening pigs, dermatitis and nephrotic syndrome (porcine dermatis and nephropathy syndrome, PDNS), young piglets diseases such as congenital tremor. In recent years, PCV2 infection in large-scale pig farms in my country and the diseases caused by it have become more and more common, causing large economic losses to pig production in my country. [0003] At present, the k...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/558G01N33/532G01N33/552C12P19/34C12N15/70C12P21/02C12R1/19
Inventor 周顺纪卫宁崔尚金李俊王金宝袁小远
Owner QINGDAO AGRI UNIV
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