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Application of Chlorine Dioxide in Prevention and Treatment of PRRS

A technology for chlorine dioxide and PRRS, which is applied in antiviral agents, active ingredients of chlorine, and pharmaceutical formulations, can solve problems such as antibody dependence enhancement, persistent infection, and short immunity period, achieving good application prospects and good Antiviral effect, no triple effect effect

Active Publication Date: 2020-01-10
广州远华生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In 2006, pig high fever broke out in my country, which caused serious economic losses to the pig industry. Later, this strain was defined as Highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV). Currently, Highly pathogenic porcine reproductive and respiratory syndrome (HP-PRRS) is one of the most damaging diseases to the pig industry. Due to the characteristics of long-term maintenance of PRRS, there are no good vaccines and drugs to prevent and control PRRS at present.
Among them, the inactivated vaccine has the following disadvantages: (1) It requires large doses of vaccination or the application of concentrated antigens, and the immunity period is short, often requiring booster vaccination; (2) It cannot cause local immunity, so that the effect of cellular immunity is weak; (3) It produces complete immunity It takes 2 to 3 weeks, which is not conducive to emergency vaccination and the reduction of vaccine costs; (4) There is a possibility of incomplete inactivation and loose virus
However, the attenuated vaccine has the risk of strong virulence, recombination and potential infection

Method used

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  • Application of Chlorine Dioxide in Prevention and Treatment of PRRS
  • Application of Chlorine Dioxide in Prevention and Treatment of PRRS
  • Application of Chlorine Dioxide in Prevention and Treatment of PRRS

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 Cytotoxicity of Chlorine Dioxide

[0045] 1. Materials

[0046] Chlorine dioxide solution (chlorine dioxide gas with a purity of 99.9% dissolved in water), provided by Shanghai Bijiasheng Technology Co., Ltd.).

[0047] AlamarBlue (purchased from Invitrogen) is used as an indicator of living cell metabolism. Under the mitochondrial enzymatic reduction reaction, a measurable fluorescent metabolite will be produced, and the cell activity can be monitored by measuring its fluorescence intensity.

[0048] 2. Test method

[0049] Cultivate Marc-145 cells or PAMs cells to 60-70% in DMEM medium containing 10% fetal bovine serum, discard the culture medium, add chlorine dioxide-containing nutrient solution for 36 hours, and set PBS control Then add 10% (v / v) ratio of AlamarBlue to continue culturing for 3 h, read the fluorescence values ​​of excitation light at 540nm and emission light at 590nm respectively with a multifunctional microplate reader, and make a chlori...

Embodiment 2

[0053] Example 2 Chlorine dioxide antiviral test with different multiplicity of infection and infection time

[0054] 1. Chlorine dioxide antiviral test with different multiplicity of infection of HP-PRRSV

[0055] (1) Western blot detection

[0056] Culture Marc-145 cells in a 6-well plate containing DMEM medium containing 10% fetal bovine serum until the cell confluence reaches 70%, discard the medium, wash with PBS three times, and inoculate HP-PRRSV with different MOI At the same time, chlorine dioxide (final concentration 36 μg / ml) was added to culture for 36 h, and a control group without chlorine dioxide was set, washed 3 times with PBS, digested with 0.25% trypsin, lysed cells, measured protein concentration, and Western-Blot.

[0057] The results are attached figure 2 Shown: Chlorine dioxide can significantly inhibit the expression of HP-PRRSV N protein, and it also has a significant inhibitory effect as the MOI of infection increases. Show that chlorine dioxide c...

Embodiment 3

[0063] Example 3 Chlorine Dioxide Pretreatment Pre-treatment Antiviral Test

[0064] Culture Marc-145 cells in 12-well plates in DMEM medium containing 10% fetal bovine serum until the cell confluency reaches 70%, discard the culture medium, wash three times with PBS, and wash with different concentrations (0, 36, 48 μg / ml) Chlorine dioxide was added to the cells and cultured for 3 h, then inoculated with HP-PRRSV at a multiplicity of infection (MOI=0.1), cultured in DMEM medium with 2% fetal bovine serum for 36 h at 37°C, washed 3 times with PBS, and collected cells Analyze on a flow cytometer or do Western-Blot to detect the antiviral effect of chlorine dioxide.

[0065] The results are attached Figure 5 Shown: Chlorine dioxide can significantly inhibit HP-PRRSV infection and N protein expression. It shows that adding chlorine dioxide first and then poisoning (pre-treatment) can significantly inhibit the infection and replication of HP-PRRSV.

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Abstract

The invention discloses application of chlorine dioxide to prevention and control of a porcine reproductive and respiratory syndrome, and in particular relates to application of high-purity chlorine dioxide liquid to preparation of a medicine for resisting a PRRSV (Porcine Reproductive and Respiratory Syndrome Virus) and / or preventing and controlling the porcine reproductive and respiratory syndrome. According to the application disclosed by the invention, experiments of each level and each phase are performed, thus proving that the chlorine dioxide still has a remarkable anti-virus effect on the porcine reproductive and respiratory syndrome virus, especially a high-pathogenicity strain HP-PRRSV for the first time, and a foundation is laid for researching a novel medicine for resisting the porcine reproductive and respiratory syndrome. The used chlorine dioxide is the high-purity chlorine dioxide liquid, is safe and non-toxic, has no mutagenic, teratogenic or carcinogenic effects and can be orally taken; the chlorine dioxide has no irritation and toxin to organisms and does not cause medicine residues, so that the chlorine dioxide is hopefully used as a novel bioactive molecule for preventing and controlling the porcine reproductive and respiratory syndrome; a prerequisite for researching and developing an anti-virus medicine for the porcine reproductive and respiratory syndrome virus by utilizing the chlorine dioxide is provided, and the chlorine dioxide has a very good application prospect in the aspects of prevention and control of the porcine reproductive and respiratory syndrome.

Description

technical field [0001] The invention belongs to the technical field of veterinary medicine. More specifically, it relates to the application of chlorine dioxide in the prevention and treatment of PRRS. Background technique [0002] Porcine reproductive and respiratory syndrome (PRRS), also known as blue ear disease, is caused by porcine reproductive and respiratory syndrome virus (Porcine reproductive and respiratory syndrome virus, PRRSV). PRRS mainly causes abortion and stillbirth in pregnant sows , mummified fetuses, weak piglets, and pigs of all ages, especially piglets, have respiratory symptoms. The characteristic lesion is interstitial pneumonia, and the mortality rate is extremely high. It is a highly contagious global important infectious disease. The disease first broke out in the United States in 1987, and then spread to Europe. The virus was isolated in my country in 1996. Currently, PRRSV is divided into two genotypes based on genome sequence and antigenicity ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K33/20A61K9/08A61P31/14
CPCA61K9/08A61K33/20
Inventor 郭春和朱振邦郭扬俞飘刘小红陈瑶生
Owner 广州远华生物科技有限公司
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