CN109453373B - Method for inactivating foot-and-mouth disease virus - Google Patents
Method for inactivating foot-and-mouth disease virus Download PDFInfo
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- CN109453373B CN109453373B CN201710796711.8A CN201710796711A CN109453373B CN 109453373 B CN109453373 B CN 109453373B CN 201710796711 A CN201710796711 A CN 201710796711A CN 109453373 B CN109453373 B CN 109453373B
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- foot
- mouth disease
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- 241000710198 Foot-and-mouth disease virus Species 0.000 title claims abstract description 47
- 238000000034 method Methods 0.000 title claims abstract description 34
- 230000000415 inactivating effect Effects 0.000 title claims description 11
- ZCCUUQDIBDJBTK-UHFFFAOYSA-N psoralen Chemical class C1=C2OC(=O)C=CC2=CC2=C1OC=C2 ZCCUUQDIBDJBTK-UHFFFAOYSA-N 0.000 claims abstract description 64
- 241000700605 Viruses Species 0.000 claims abstract description 45
- VXGRJERITKFWPL-UHFFFAOYSA-N 4',5'-Dihydropsoralen Natural products C1=C2OC(=O)C=CC2=CC2=C1OCC2 VXGRJERITKFWPL-UHFFFAOYSA-N 0.000 claims abstract description 30
- 238000004113 cell culture Methods 0.000 claims abstract description 29
- 238000005286 illumination Methods 0.000 claims abstract description 10
- -1 psoralen compound Chemical class 0.000 claims description 9
- 230000035515 penetration Effects 0.000 claims description 3
- 230000002779 inactivation Effects 0.000 abstract description 29
- 229960005486 vaccine Drugs 0.000 abstract description 10
- 239000000427 antigen Substances 0.000 abstract description 6
- 102000036639 antigens Human genes 0.000 abstract description 6
- 108091007433 antigens Proteins 0.000 abstract description 6
- 241001465754 Metazoa Species 0.000 abstract description 5
- 230000000694 effects Effects 0.000 abstract description 3
- 230000000937 inactivator Effects 0.000 abstract description 3
- 241000894006 Bacteria Species 0.000 abstract description 2
- 230000009286 beneficial effect Effects 0.000 abstract description 2
- 238000011109 contamination Methods 0.000 abstract description 2
- 230000002349 favourable effect Effects 0.000 abstract description 2
- 231100000252 nontoxic Toxicity 0.000 abstract description 2
- 230000003000 nontoxic effect Effects 0.000 abstract description 2
- 238000003860 storage Methods 0.000 abstract description 2
- 208000007212 Foot-and-Mouth Disease Diseases 0.000 description 9
- 239000012930 cell culture fluid Substances 0.000 description 6
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 4
- 229930006000 Sucrose Natural products 0.000 description 4
- 238000003556 assay Methods 0.000 description 4
- 239000005720 sucrose Substances 0.000 description 4
- 229940027041 8-mop Drugs 0.000 description 3
- QXKHYNVANLEOEG-UHFFFAOYSA-N Methoxsalen Chemical compound C1=CC(=O)OC2=C1C=C1C=COC1=C2OC QXKHYNVANLEOEG-UHFFFAOYSA-N 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 3
- 229940031551 inactivated vaccine Drugs 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 102000035195 Peptidases Human genes 0.000 description 2
- 108091005804 Peptidases Proteins 0.000 description 2
- 241000709664 Picornaviridae Species 0.000 description 2
- GLUUGHFHXGJENI-UHFFFAOYSA-N Piperazine Chemical compound C1CNCCN1 GLUUGHFHXGJENI-UHFFFAOYSA-N 0.000 description 2
- 239000004365 Protease Substances 0.000 description 2
- 230000015556 catabolic process Effects 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 238000006731 degradation reaction Methods 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 244000052769 pathogen Species 0.000 description 2
- 241000283690 Bos taurus Species 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 241000282887 Suidae Species 0.000 description 1
- 241001493546 Suina Species 0.000 description 1
- 208000003441 Transfusion reaction Diseases 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 230000001464 adherent effect Effects 0.000 description 1
- 239000012503 blood component Substances 0.000 description 1
- 210000000234 capsid Anatomy 0.000 description 1
- 238000004132 cross linking Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000001627 detrimental effect Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000010494 dissociation reaction Methods 0.000 description 1
- 230000005593 dissociations Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 239000012510 hollow fiber Substances 0.000 description 1
- 230000003053 immunization Effects 0.000 description 1
- 238000002649 immunization Methods 0.000 description 1
- 230000002163 immunogen Effects 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000002458 infectious effect Effects 0.000 description 1
- 210000000265 leukocyte Anatomy 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 238000001471 micro-filtration Methods 0.000 description 1
- 244000000010 microbial pathogen Species 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 238000004886 process control Methods 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000004114 suspension culture Methods 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
- A61K41/10—Inactivation or decontamination of a medicinal preparation prior to administration to an animal or a person
- A61K41/17—Inactivation or decontamination of a medicinal preparation prior to administration to an animal or a person by ultraviolet [UV] or infrared [IR] light, X-rays or gamma rays
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/12—Viral antigens
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
- A61K2039/525—Virus
- A61K2039/5252—Virus inactivated (killed)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/55—Medicinal preparations containing antigens or antibodies characterised by the host/recipient, e.g. newborn with maternal antibodies
- A61K2039/552—Veterinary vaccine
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
- C12N2770/00011—Details
- C12N2770/32011—Picornaviridae
- C12N2770/32111—Aphthovirus, e.g. footandmouth disease virus
- C12N2770/32134—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Virology (AREA)
- Immunology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
The invention provides an inactivation method of foot-and-mouth disease virus, which comprises the following steps: adding psoralen compounds into the foot-and-mouth disease virus cell culture solution, and then carrying out illumination treatment to obtain the inactivated foot-and-mouth disease virus solution. The invention has the following beneficial effects: 1. the inactivation process is greatly simplified, and the inactivation process time is shortened; 2. the stability of the inactivation process is improved; 3. the loss of antigen is reduced; 4. compared with other inactivators, psoralen is nontoxic and does not cause immune side effects on animals; 5. has the functions of eliminating other bacteria and viruses, and is favorable to raising the contamination resistance of vaccine, raising product quality and prolonging storage time.
Description
Technical Field
The invention relates to the technical field of foot-and-mouth disease virus inactivation, in particular to a method for inactivating foot-and-mouth disease virus.
Background
Foot-and-mouth disease (FMD) is an acute, hot, highly contagious and rapidly long-distance transmissible animal epidemic disease caused by foot-and-mouth disease virus (FMDV), the infected animals are main livestock species such as pigs, cows and sheep and other domestic and wild artiodactyls, and the susceptible animals are more than 70. At present, vaccine immunization is one of the most important and successful prevention measures for foot-and-mouth disease, wherein the conventional inactivated vaccine is the most main vaccine for the current foot-and-mouth disease immune control.
Inactivated vaccines are vaccines that are made by inactivating infectious pathogens by a suitable physical or chemical means such that the pathogens are rendered non-pathogenic, but remain immunogenic. Therefore, the selection of proper inactivator to inactivate the foot-and-mouth disease virus is an important link for manufacturing inactivated vaccines. The inactivation process of the foot-and-mouth disease virus adopts the inactivation mode of diethylene imine (BEI) and beta-propyl ester at present, and the method is widely applied to production but causes higher antigen loss, long inactivation time and complex process control, and is a key factor causing low production efficiency of the current foot-and-mouth disease inactivated virus vaccine.
Psoralen is also called furocoumarin and is widely applied to blood disinfection at present, which can not only inactivate known and unknown pathogenic microorganisms in blood, but also remove leukocytes and cytokines in single blood components such as plasma and platelets, thereby avoiding the occurrence of blood transfusion reaction. However, picornaviruses are resistant to the inactivation of psoralens because the viral capsid is very dense and psoralens are difficult to pass through.
Foot-and-mouth disease virus is used as a picornavirus, and under the conventional reaction condition, psoralen molecules cannot enter the interior of virus particles so as to photocrosslink RNA to inactivate the virus; and foot-and-mouth disease virus is sensitive and unstable to environmental conditions, and the conventional inactivation condition easily causes antigen loss. Therefore, no report of foot-and-mouth disease virus inactivation by psoralen exists at present.
Disclosure of Invention
Aiming at the defects in the prior art, the invention provides a method for inactivating foot-and-mouth disease virus.
The purpose of the invention is realized by the following technical scheme:
the invention provides an inactivation method of foot-and-mouth disease virus, which comprises the following steps: adding psoralen compounds into the foot-and-mouth disease virus cell culture solution, and then carrying out illumination treatment to obtain the inactivated foot-and-mouth disease virus solution.
Preferably, the electrical conductivity of the virus cell culture solution system is 0.5-15 mS/cm; more preferably, the conductance is 1-10 mS/cm. The virus cell culture fluid has low electrical conductance, which can cause virus dissociation; too high a conductivity can result in the failure of the psoralen molecule to enter.
Preferably, after the psoralen compounds are added into the virus cell culture solution, the temperature of the virus cell culture solution is controlled to be 26-38 ℃ during the ultraviolet illumination period. Too high a temperature and too long a treatment time can lead to degradation of the virus by proteases, while too low a temperature is detrimental to the entry of psoralen molecules into the virus.
Preferably, the molar concentration of the added psoralen compound is 100-20000 times that of the foot-and-mouth disease virus.
More preferably, the molar concentration of the added psoralen compound is 200-10000 times of that of the foot-and-mouth disease virus. The concentration of the psoralen is 1-100 mg/L. When the psoralen molecules are in the concentration ratio range, absolute excess of relative virus concentration can be ensured, the inactivation reaction is facilitated, and the reduction of relative concentration caused by adsorption and nonspecific crosslinking of other impurity molecules on the psoralen molecules and the virus inactivation are also prevented.
Preferably, the psoralen compound is psoralen and derivatives thereof, specifically including AMT, S-59, TMP, and 8-MOP.
More preferably, the psoralen compound is AMT or S-59.
Preferably, the parameters specifically adopted for the illumination treatment include: the light wavelength is 300-400nm, more preferably 340-380nm, and the irradiation dose of the ultraviolet illumination is 3-8mW/cm2The treatment time is 10-100 minutes.
Preferably, the light treatment is carried out by passing the virus cell culture fluid added with psoralen through a long and narrow tube having ultraviolet ray penetration.
The inactivation method is not only suitable for foot-and-mouth disease virus, but also suitable for inactivation of other viruses, and can achieve the effect of the invention.
Compared with the prior art, the invention has the following beneficial effects:
1. the inactivation process is greatly simplified, and the inactivation process time is shortened;
2. the stability of the inactivation process is improved;
3. the loss of antigen is reduced;
4. compared with other inactivators, the psoralen is nontoxic and does not cause immune side effects on animals;
5. has the functions of eliminating other bacteria and viruses, and is favorable to raising the contamination resistance of vaccine, raising product quality and prolonging storage time.
Detailed Description
The present invention will be described in detail with reference to specific examples. The following examples will assist those skilled in the art in further understanding the invention, but are not intended to limit the invention in any way. It should be noted that variations and modifications can be made by persons skilled in the art without departing from the spirit of the invention. All falling within the scope of the present invention.
The embodiment of the invention provides a method for inactivating foot-and-mouth disease virus, which comprises the following steps: adding psoralen compounds into the foot-and-mouth disease virus cell culture solution, and then carrying out illumination treatment to obtain the inactivated foot-and-mouth disease virus solution.
The conductance of the virus cell culture solution system is 0.5-15 mS/cm; more preferably, the conductance is 1-10 mS/cm.
After the psoralen compounds are added into the virus cell culture solution, the temperature of the virus cell culture solution is controlled to be 26-38 ℃ during ultraviolet illumination. Too high a temperature and too long a treatment time can lead to the degradation of the virus by proteases, while too low a temperature is not conducive to the entry of psoralen molecules into the interior of the virus molecules.
The molar concentration of the added psoralen compounds is 100-20000 times of that of the foot-and-mouth disease virus. More preferably 200-10000 times.
The psoralen compounds are psoralen and its derivatives, specifically including AMT, S-59, TMP, and 8-MOP.
The psoralen compounds are AMT and S-59.
The parameters specifically adopted for the illumination treatment include: the light wavelength is 340-2The treatment time is 10-100 minutes.
The light treatment is carried out by passing the virus cell culture fluid added with psoralen through a long and narrow pipeline with ultraviolet penetration.
Example 1
The embodiment provides an inactivation method of a foot-and-mouth disease vaccine, which adopts psoralen photochemistry to carry out scale virus antigen inactivation and comprises the following specific steps:
(1) filtering the A-type foot-and-mouth disease virus cell culture solution obtained by suspension culture of the BHK21 cells through a 0.22 mu m microfiltration membrane to obtain a clarified virus cell culture solution.
(2) The virus cell culture fluid is concentrated by 10 times through a 300kD hollow fiber membrane, and the conductance of the virus cell culture fluid is reduced to 1mS/cm by washing and filtering the virus cell culture fluid with a PB solution.
(3) Psoralen S-59 was added to the clarified virus cell culture to a final concentration of 0.625 mL/L. The molar concentration of S-59 is 200 times of that of the foot-and-mouth disease virus.
(4) Passing virus cell culture solution containing psoralen through transparent tube, and applying ultraviolet light with illuminance of 8mW/cm2Ultraviolet rays are applied to the virus cell culture solution flowing through the transparent tube, and the residence time under the ultraviolet rays is kept for 10 minutes while the temperature is maintained at 38 ℃.
(5) The virus cell culture solution irradiated with ultraviolet light by the method of this example was tested for TCID50 to be 0 by plaque assay; the content of 146S in the virus cell culture solution before and after inactivation is respectively 51.31 mu g/mL and 50.29 mu g/mL by the sucrose density gradient method.
Example 2
The embodiment provides an inactivation method of a foot-and-mouth disease vaccine, which adopts psoralen photochemistry to carry out scale virus antigen inactivation and comprises the following specific steps:
(1) a culture solution of O-type foot-and-mouth disease virus cells cultured by BHK21 adherent cell shaking flasks is centrifuged at 6000g for 15 minutes at 4 ℃ to obtain a clear virus cell culture solution.
(2) The solution conductance was adjusted to 10mS/cm using PB solution.
(3) Psoralen AMT was added to the clarified virus cell culture to a final concentration of 5 mg/L. The molar concentration of the AMT is 20000 times of that of the foot-and-mouth disease virus.
(4) Subjecting the virus cell culture solution containing psoralen to ultraviolet irradiation of 3mW/cm2Ultraviolet irradiation was performed while maintaining the temperature at 26 ℃ for 100 minutes.
(5) The virus cell culture solution irradiated with ultraviolet light by the method of this example was 0 in TCID50 measured by plaque assay; the content of 146S in the virus cell culture solution before and after inactivation is respectively 4.33 mug/mL and 4.31 mug/mL by the sucrose density gradient method.
Example 3
The embodiment provides an inactivation method of a foot-and-mouth disease vaccine, which has the specific steps basically the same as those of the embodiment 2, and the difference is only that: the psoralen compound used in this example was 8-MOP.
The virus cell culture solution irradiated with ultraviolet light by the method of this example had TCID50 of 10 to 1.5 as measured by plaque assay; the content of 146S in the virus cell culture solution before and after inactivation is respectively 3.35 mug/mL and 2.1 mug/mL by the sucrose density gradient method.
Example 4
The embodiment provides an inactivation method of a foot-and-mouth disease vaccine, which has the specific steps basically the same as those of the embodiment 1, and the difference is only that: step 2 employed in this example was to adjust the conductivity of the solution to 0.5mS/cm using PB solution.
The virus cell culture solution irradiated with ultraviolet light by the method of this example was found to have TCID50 not 0 by plaque assay; the content of 146S in the virus cell culture solution before and after inactivation is respectively 22.30 mu g/mL and 20.04 mu g/mL by the sucrose density gradient method.
The specific application of the invention is numerous and the above description is only a preferred embodiment of the invention. It should be noted that the above examples are only for illustrating the present invention, and are not intended to limit the scope of the present invention. It will be apparent to those skilled in the art that various modifications can be made without departing from the principles of the invention and these modifications are to be considered within the scope of the invention.
Claims (6)
1. The method for inactivating the foot-and-mouth disease virus is characterized by comprising the following steps of: adding psoralen compounds into the foot-and-mouth disease virus cell culture solution, and then carrying out illumination treatment to obtain an inactivated foot-and-mouth disease virus solution;
the psoralen compounds are AMT and S-59;
the conductance of the virus cell culture solution is 1-15 mS/cm.
2. The method for inactivating foot and mouth disease virus according to claim 1, wherein the temperature of the culture solution is controlled to be 26-38 ℃ after the psoralen compound is added to the virus cell culture solution.
3. The method for inactivating foot and mouth disease virus according to claim 1, wherein the molar concentration of the psoralen compound is 100-20000 times that of the foot and mouth disease virus.
4. The method for inactivating foot and mouth disease virus according to claim 3, wherein the molar concentration of the psoralen compound is 200-10000 times that of the foot and mouth disease virus.
5. The method of claim 1, wherein the parameters specifically adopted for the light treatment include: the light wavelength is 300-400nm, and the ultraviolet illumination is 3-8mW/cm2The treatment time is 10-100 minutes.
6. The method for inactivating foot and mouth disease virus according to claim 1, wherein the light treatment is carried out by passing a virus cell culture solution added with psoralen through a long and narrow tube having ultraviolet ray penetration.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201710796711.8A CN109453373B (en) | 2017-09-06 | 2017-09-06 | Method for inactivating foot-and-mouth disease virus |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201710796711.8A CN109453373B (en) | 2017-09-06 | 2017-09-06 | Method for inactivating foot-and-mouth disease virus |
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| CN109453373A CN109453373A (en) | 2019-03-12 |
| CN109453373B true CN109453373B (en) | 2021-08-17 |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6004742A (en) * | 1993-06-28 | 1999-12-21 | Cerus Corporation | Method for inactivation of pathogens in platelets using 4' and 5' primary amino-substituted psoralens |
| CN1566114A (en) * | 2003-06-16 | 2005-01-19 | 高和英 | Novel psoralen derivative for annihilating live virus or germ |
| CN1857240A (en) * | 2006-04-06 | 2006-11-08 | 中国农业科学院兰州畜牧与兽药研究所 | Application of hypericin in preparing RNA virus resisting medicine |
| CN102933211A (en) * | 2009-12-21 | 2013-02-13 | 佐治亚州立大学研究基金会 | Photo-inactivated viruses and systems and methods of using same |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2001055354A1 (en) * | 2000-01-31 | 2001-08-02 | Consortium For Plasma Science, Llc | Methods for inactivating viruses |
| CN104311673B (en) * | 2014-10-22 | 2017-04-12 | 申联生物医药（上海）股份有限公司 | Method for preparing pig O-type foot-and-mouth disease synthetic peptide antigen 2800 by using solid-phase fragment method |
-
2017
- 2017-09-06 CN CN201710796711.8A patent/CN109453373B/en active Active
Patent Citations (4)
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|---|---|---|---|---|
| US6004742A (en) * | 1993-06-28 | 1999-12-21 | Cerus Corporation | Method for inactivation of pathogens in platelets using 4' and 5' primary amino-substituted psoralens |
| CN1566114A (en) * | 2003-06-16 | 2005-01-19 | 高和英 | Novel psoralen derivative for annihilating live virus or germ |
| CN1857240A (en) * | 2006-04-06 | 2006-11-08 | 中国农业科学院兰州畜牧与兽药研究所 | Application of hypericin in preparing RNA virus resisting medicine |
| CN102933211A (en) * | 2009-12-21 | 2013-02-13 | 佐治亚州立大学研究基金会 | Photo-inactivated viruses and systems and methods of using same |
Non-Patent Citations (2)
| Title |
|---|
| Olga Ferna＇ndez-Miragall,et al..In vivo footprint of a picornavirus internal ribosome entry site reveals differences in accessibility to specific RNA structural elements.《Journal of General Virology》.2007,第88卷第3053-3062页. * |
| 补骨脂素光化学法在血液消毒方面的研究进展;饶林等;《中国消毒学杂志》;20031231;第20卷(第2期);第147-150页 * |
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