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Colloidal gold quick diagnosis test paper for distinguishing classic porcine reproductive and respiratory syndrome (PRRS) from highly pathogenic PRRS (HPRRS)

A rapid diagnosis, colloidal gold technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the limitations of the prevention and control of porcine PRRS, high detection costs, inconvenient real-time and rapid differential diagnosis of classic porcine PRRS and high pathogenicity To solve the problems of pig blue-ear disease and other problems, achieve the effect of low production cost, low detection cost, and improve sensitivity and specificity

Inactive Publication Date: 2012-08-01
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Faster and more accurate techniques are ELISA and fluorescent RT-PCR, but these two techniques require special equipment, and the detection time is more than 2 hours, the detection cost is expensive, and it is not convenient for real-time and rapid differential diagnosis of classic PRRS and highly pathogenic porcine blue ear disease
These limit the prevention and control of pig PRRS, so it is imminent to develop a diagnostic method with strong specificity, high sensitivity and simple operation

Method used

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  • Colloidal gold quick diagnosis test paper for distinguishing classic porcine reproductive and respiratory syndrome (PRRS) from highly pathogenic PRRS (HPRRS)
  • Colloidal gold quick diagnosis test paper for distinguishing classic porcine reproductive and respiratory syndrome (PRRS) from highly pathogenic PRRS (HPRRS)
  • Colloidal gold quick diagnosis test paper for distinguishing classic porcine reproductive and respiratory syndrome (PRRS) from highly pathogenic PRRS (HPRRS)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1 Preparation of Immunogen

[0020] The deletion of 30 amino acids in Nsp2 has become an important index for distinguishing highly pathogenic PRRS from classical PRRS. In this experiment, 30 amino acids of the PRRS Nsp2 region as an antigen were synthesized by the company's synthetic method and coupled with the BSA carrier to prepare the immunogen.

Embodiment 2

[0021] Example 2 Preparation of anti-classic PRRS monoclonal antibody

[0022] 1. For animal immunization, select healthy Balb / c mice aged 6-8 weeks, and inject about 50 μg of the immunogen emulsified with Freund's complete adjuvant into the abdominal cavity of each mouse, and emulsify the immunogen protein with Freund's incomplete adjuvant 14 days later 100 μg was injected intraperitoneally, and 100 μg of purified immunogenic protein was injected intraperitoneally directly at the last booster immunization, and 50 μg of purified immunogenic protein was injected into the tail vein 3 to 4 days before fusion.

[0023] 2. Cell fusion Take the splenocytes of immunized mice and mix them with SP2 / 0 in the fusion tube, centrifuge at 300g for 10min, discard the supernatant, shake the cells to mix the two kinds of cells as evenly as possible, and then add the preheated solution slowly within 45s. PEG solution, then slowly add serum-free 1640 medium to stop the fusion, let stand and th...

Embodiment 3

[0032] Example 3 Preparation of colloidal gold and gold-labeled antigen

[0033] 1. Preparation of colloidal gold:

[0034] Dilute 1% chloroauric acid to 0.01% with double-distilled deionized water, heat to boil, add 2ml 1% sodium citrate for every 100ml 0.01% chloroauric acid, continue to boil until the liquid is bright red, then stop heating, cool to Rehydrate at room temperature. The appearance of the prepared colloidal gold should be pure, translucent, free of precipitation and floating matter.

[0035] 2. Preparation of colloidal gold-labeled classic PRRS specific antigen:

[0036] Use 0.1M potassium carbonate to adjust the pH value of the colloidal gold to 8.2, add the classic PRRS specific antigen at 100ug antibody / ml colloidal gold, mix with a magnetic stirrer for 30 minutes, add BSA to a final concentration of 1%, and let it stand for 1 hour. Centrifuge at 13000rpm at 4°C for 30 minutes, discard the supernatant, wash the precipitate twice with the labeled washing...

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Abstract

The invention relates to colloidal gold quick diagnosis test paper for distinguishing classic porcine reproductive and respiratory syndrome (PRRS) from highly pathogenic PRRS (HPRRS). The test paper is characterized in that a sample pad, a bonding pad, a nitrocellulose membrane and an absorption pad are sequentially adhered to a nonabsorbent support sheet; the bonding pad is coated with a colloidal gold labeled classic PRRS virus (PRRSV) peculiar antigen; and the nitrocellulose membrane is respectively coated with a detection line, namely a T line consisting of staphylococcal protein A (SPA), and a quality control line, namely a C line consisting of a classic PRRS monoclonal antibody 2B9. The test paper has the obvious advantages of high specificity, sensitivity and diagnosis speed, is easy to operate, can be used for distinguishing classic PRRSV from HPRRSV, and is used for the quick diagnosis of PRRSV.

Description

technical field [0001] The invention relates to a colloidal gold rapid diagnostic test strip for distinguishing classic PRRS and HPRRS, which belongs to a new rapid and specific diagnostic method, and is mainly used for clinical porcine reproductive and respiratory syndrome (commonly known as blue ear disease, PRRS) and high Differential diagnosis and epidemiological investigation of pathogenic porcine blue-ear disease (HPRRS); applied in the field of animal husbandry and veterinary medicine. Background technique [0002] Porcine reproductive and respiratory syndrome (PRRS) is a reproductive disorder in pregnant sows caused by porcine reproductive and respiratory syndrome virus (PRRSV). An infectious disease characterized by respiratory symptoms in piglets. The disease first broke out in the United States in 1987, and then quickly spread to all parts of the world. It has become one of the important diseases that endanger large-scale pig production. Since 2001, some areas o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/569
Inventor 丁壮刘慧母连志韩明明姜翠翠李想朱利塞黄志强
Owner JILIN UNIV
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