Inactivation of Poliovirus by Ozone and the Impact of Ozone on the Viral Genome
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摘要
Objective To investigate the mechanisms underlying ozone‐induced inactivation of poliovirus type 1(PV1). Methods We used cell culture, long‐overlapping RT‐PCR, and spot hybridization assays to verify and accurately locate the sites of action of ozone that cause PV1 inactivation. We also employed recombinant viral genome RNA infection models to confirm our observations. Results Our results indicated that ozone inactivated PV1 primarily by disrupting the 5′‐non‐coding region(5′‐NCR) of the PV1 genome. Further study revealed that ozone specifically damaged the 80‐124 nucleotide(nt) region in the 5′‐NCR. Recombinant viral genome RNA infection models confirmed that PV1 lacking this region was non‐infectious. Conclusion In this study, we not only elucidated the mechanisms by which ozone induces PV1 inactivation but also determined that the 80‐124 nt region in the 5′‐NCR is targeted by ozone to achieve this inactivation.
Objective To investigate the mechanisms underlying ozone‐induced inactivation of poliovirus type 1(PV1). Methods We used cell culture, long‐overlapping RT‐PCR, and spot hybridization assays to verify and accurately locate the sites of action of ozone that cause PV1 inactivation. We also employed recombinant viral genome RNA infection models to confirm our observations. Results Our results indicated that ozone inactivated PV1 primarily by disrupting the 5′‐non‐coding region(5′‐NCR) of the PV1 genome. Further study revealed that ozone specifically damaged the 80‐124 nucleotide(nt) region in the 5′‐NCR. Recombinant viral genome RNA infection models confirmed that PV1 lacking this region was non‐infectious. Conclusion In this study, we not only elucidated the mechanisms by which ozone induces PV1 inactivation but also determined that the 80‐124 nt region in the 5′‐NCR is targeted by ozone to achieve this inactivation.
Objective To investigate the mechanisms underlying ozone‐induced inactivation of poliovirus type 1(PV1). Methods We used cell culture, long‐overlapping RT‐PCR, and spot hybridization assays to verify and accurately locate the sites of action of ozone that cause PV1 inactivation. We also employed recombinant viral genome RNA infection models to confirm our observations. Results Our results indicated that ozone inactivated PV1 primarily by disrupting the 5′‐non‐coding region(5′‐NCR) of the PV1 genome. Further study revealed that ozone specifically damaged the 80‐124 nucleotide(nt) region in the 5′‐NCR. Recombinant viral genome RNA infection models confirmed that PV1 lacking this region was non‐infectious. Conclusion In this study, we not only elucidated the mechanisms by which ozone induces PV1 inactivation but also determined that the 80‐124 nt region in the 5′‐NCR is targeted by ozone to achieve this inactivation.
引文
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2.Alam MM,Sharif S,Shaukat S,et al.Genomic Surveillance Elucidates Persistent Wild Poliovirus Transmission During2013‐2015 in Major Reservoir Areas of Pakistan.Clin Infect Dis,2016;62,190‐8.
3.Famulare M.Has Wild Poliovirus Been Eliminated from Nigeria.PLoS One,2015;10,e0135765.
4.Moran‐Gilad J,Kaliner E,Gdalevich M,et al.Public health response to the silent reintroduction of wild poliovirus to Israel,2013‐2014.Clin Microbiol Infect,2016;22,S140‐S145.
5.Jin M,Shan J,Chen Z,et al.Chlorine dioxide inactivation of enterovirus 71 in water and its impact on genomic targets.Environ Sci Technol,2013;47,4590‐7.
6.Lukasik J,Bradley ML,Scott TM,et al.Reduction of poliovirus 1,bacteriophages,Salmonella montevideo,and Escherichia coli O157:H7 on strawberries by physical and disinfectant washes.JFood Prot,2003;66,188‐93.
7.Tree JA,Adams MR,Lees DN.Chlorination of indicator bacteria and viruses in primary sewage effluent.Appl Environ Microbiol,2003;69,2038‐43.
8.Nuanualsuwan S,Mariam T,Himathongkham S,et al.Ultraviolet inactivation of feline calicivirus,human enteric viruses and coliphages.Photochem Photobiol,2002;76,406‐10.
9.Apaire‐Marchais V,Robertson BH,Aubineau‐Ferre V,et al.Direct sequencing of hepatitis A virus strains isolated during an epidemic in France.Appl Environ Microbiol,1995;61,3977‐80.
10.Churn CC,Bates RC,Boardman GD.Mechanism of chlorine inactivation of DNA‐containing parvovirus H‐1.Appl Environ Microbiol,1983;46,1394‐402.
11.LYK N,RMB F,VRDS F,et al.Pre‐ozonation of source water:Assessment of efficacy against Giardia duodenalis cysts and effects on natural organic matter.Chemosphere,2019;214,764‐70.
12.von GU.Oxidation Processes in Water Treatment:Are We on Track.Environ Sci Technol,2018;52,5062‐75.
13.Dong S,Massalha N,Plewa MJ,et al.The impact of disinfection Ct values on cytotoxicity of agricultural wastewaters:Ozonation vs.chlorination.Water Res,2018;144,482‐90.
14.Wang H,Hu C,Zhang S,et al.Effects of O3/Cl2 disinfection on corrosion and opportunistic pathogens growth in drinking water distribution systems.J Environ Sci,2018;73,38‐46.
15.?nnby L,Salhi E,McKay G,et al.Ozone and chlorine reactions with dissolved organic matter‐Assessment of oxidant‐reactive moieties by optical measurements and the electron donating capacities.Water Res,2018;144,64‐75.
16.Tamaki M,Kobayashi F,Ikeura H,et al.Disinfection by Ozone Microbubbles Can Cause Morphological Change of Fusarium oxysporum f.sp.melonis Spores.Plant Pathol J,2018;34,335‐40.
17.Ofori I,Maddila S,Lin J,et al.Ozone initiated inactivation of Escherichia coli and Staphylococcus aureus in water:Influence of selected organic solvents prevalent in wastewaters.Chemosphere,2018;206,43‐50.
18.Roy D,Wong PK,Engelbrecht RS,et al.Mechanism of enteroviral inactivation by ozone.Appl Environ Microbiol,1981;41,718‐23.
19.Kitamura N.[Genome structure of poliovirus(author's transl)].Seikagaku,1982;54,151‐6.
20.Simonet J,Gantzer C.Inactivation of poliovirus 1 and F‐specific RNA phages and degradation of their genomes by UVirradiation at 254 nanometers.Appl Environ Microbiol,2006;72,7671‐7.
21.Bhattacharya SS,Kulka M,Lampel KA,et al.Use of reverse transcription and PCR to discriminate between infectious and non‐infectious hepatitis A virus.J Virol Methods,2004;116,181‐7.
22.Jin M,Zhao ZG,Wang XW,et al.The 40‐80?nt region in the5'‐NCR of genome is a critical target for inactivating poliovirus by chlorine dioxide.J Med Virol,2012;84,526‐35.
23.Zaidi SS,Asghar H,Sharif S,et al.Poliovirus Laboratory Based Surveillance:An Overview.Methods Mol Biol,2016;1387,11‐8.
24.Katzenelson E,Biedermann N.Disinfection of viruses in sewage by ozone.Water Res,1976;10,629‐31.
25.Chatterjee NK,Kaehler M,Deibel R.Detection of enteroviruses using subgenomic probes of Coxsackie virus B4 by hybridization.Diagn Microbiol Infect Dis,1988;11,129‐36.
26.Cello J,Paul AV,Wimmer E.Chemical synthesis of poliovirus cDNA:generation of infectious virus in the absence of natural template.Science,2002;297,1016‐8.
27.Nuanualsuwan S,Cliver DO.Infectivity of RNA from inactivated poliovirus.Appl Environ Microbiol,2003;69,1629‐32.
28.Nomoto A,Omata T,Toyoda H,et al.Complete nucleotide sequence of the attenuated poliovirus Sabin 1 strain genome.Proc Natl Acad Sci USA,1982;79,5793‐7.
29.Camel V,Bermond A.The use of ozone and associated oxidation processes in drinking water treatment.Water Res,1998;32,3208‐22.
30.Moore NJ,Margolin AB.Efficacy of Nucleic Acid Probes for Detection of Poliovirus in Water Disinfected by Chlorine,Chlorine Dioxide,Ozone,and UV Radiation.Appl Environ Microbiol,1994;60,4189‐91.
31.Simonet J,Gantzer C.Degradation of the Poliovirus 1 genome by chlorine dioxide.J Appl Microbiol,2006;100,862‐70.
32.Li JW,Xin ZT,Wang XW,et al.Mechanisms of inactivation of hepatitis A virus in water by chlorine dioxide.Water Res,2004;38,1514‐9.
33.Souii A,Ben MM,Gharbi J.Role of RNA structure motifs in IRES‐dependent translation initiation of the coxsackievirus B3:new insights for developing live‐attenuated strains for vaccines and gene therapy.Mol Biotechnol,2013;55,179‐202.
34.Moustafa IM,Gohara DW,Uchida A,et al.Conformational Ensemble of the Poliovirus 3CD Precursor Observed by MDSimulations and Confirmed by SAXS:A Strategy to Expand the Viral Proteome.Viruses,2015;7,5962‐86.
35.Harris KS,Xiang W,Alexander L,et al.Interaction of poliovirus polypeptide 3CDpro with the 5'and 3'termini of the poliovirus genome.Identification of viral and cellular cofactors needed for efficient binding.J Biol Chem,1994;269,27004‐14.
36.Ogram SA,Spear A,Sharma N,et al.The 5'CL‐PCBP RNPcomplex,3'poly(A)tail and 2A(pro)are required for optimal translation of poliovirus RNA.Virology,2010;397,14‐22.
37.Schmid M,Wimmer E.IRES‐controlled protein synthesis and genome replication of poliovirus.Arch Virol Suppl,1994;9,279‐89.
38.Fitzgerald KD,Semler BL.Re‐localization of cellular protein SRp20 during poliovirus infection:bridging a viral IRES to the host cell translation apparatus.PLo S Pathog,2011;7,e1002127.
39.Muslin C,Joffret ML,Pelletier I,et al.Evolution and Emergence of Enteroviruses through Intra‐and Inter‐species Recombination:Plasticity and Phenotypic Impact of Modular Genetic Exchanges in the 5'Untranslated Region.PLoS Pathog,2015;11,e1005266.
40.Viktorova EG,Khattar S,Samal S,et al.Poliovirus Replicon RNAGeneration,Transfection,Packaging,and Quantitation of Replication.Curr Protoc Microbiol,2018;48,15H.4.1‐15H.4.15.
41.Ma JF,Straub TM,Pepper IL,et al.Cell culture and PCRdetermination of poliovirus inactivation by disinfectants.Appl Environ Microbiol,1994;60,4203‐6.
42.Maier,Tougianidou,Wiedenmann,et al.Detection of poliovirus by cell culture and by PCR after UV disinfection.Water Sci Technoly,1995;31,141‐5.
43.Lim MY,Kim JM,Ko G.Disinfection kinetics of murine norovirus using chlorine and chlorine dioxide.Water Res,2010;44,3243‐51.
2.Alam MM,Sharif S,Shaukat S,et al.Genomic Surveillance Elucidates Persistent Wild Poliovirus Transmission During2013‐2015 in Major Reservoir Areas of Pakistan.Clin Infect Dis,2016;62,190‐8.
3.Famulare M.Has Wild Poliovirus Been Eliminated from Nigeria.PLoS One,2015;10,e0135765.
4.Moran‐Gilad J,Kaliner E,Gdalevich M,et al.Public health response to the silent reintroduction of wild poliovirus to Israel,2013‐2014.Clin Microbiol Infect,2016;22,S140‐S145.
5.Jin M,Shan J,Chen Z,et al.Chlorine dioxide inactivation of enterovirus 71 in water and its impact on genomic targets.Environ Sci Technol,2013;47,4590‐7.
6.Lukasik J,Bradley ML,Scott TM,et al.Reduction of poliovirus 1,bacteriophages,Salmonella montevideo,and Escherichia coli O157:H7 on strawberries by physical and disinfectant washes.JFood Prot,2003;66,188‐93.
7.Tree JA,Adams MR,Lees DN.Chlorination of indicator bacteria and viruses in primary sewage effluent.Appl Environ Microbiol,2003;69,2038‐43.
8.Nuanualsuwan S,Mariam T,Himathongkham S,et al.Ultraviolet inactivation of feline calicivirus,human enteric viruses and coliphages.Photochem Photobiol,2002;76,406‐10.
9.Apaire‐Marchais V,Robertson BH,Aubineau‐Ferre V,et al.Direct sequencing of hepatitis A virus strains isolated during an epidemic in France.Appl Environ Microbiol,1995;61,3977‐80.
10.Churn CC,Bates RC,Boardman GD.Mechanism of chlorine inactivation of DNA‐containing parvovirus H‐1.Appl Environ Microbiol,1983;46,1394‐402.
11.LYK N,RMB F,VRDS F,et al.Pre‐ozonation of source water:Assessment of efficacy against Giardia duodenalis cysts and effects on natural organic matter.Chemosphere,2019;214,764‐70.
12.von GU.Oxidation Processes in Water Treatment:Are We on Track.Environ Sci Technol,2018;52,5062‐75.
13.Dong S,Massalha N,Plewa MJ,et al.The impact of disinfection Ct values on cytotoxicity of agricultural wastewaters:Ozonation vs.chlorination.Water Res,2018;144,482‐90.
14.Wang H,Hu C,Zhang S,et al.Effects of O3/Cl2 disinfection on corrosion and opportunistic pathogens growth in drinking water distribution systems.J Environ Sci,2018;73,38‐46.
15.?nnby L,Salhi E,McKay G,et al.Ozone and chlorine reactions with dissolved organic matter‐Assessment of oxidant‐reactive moieties by optical measurements and the electron donating capacities.Water Res,2018;144,64‐75.
16.Tamaki M,Kobayashi F,Ikeura H,et al.Disinfection by Ozone Microbubbles Can Cause Morphological Change of Fusarium oxysporum f.sp.melonis Spores.Plant Pathol J,2018;34,335‐40.
17.Ofori I,Maddila S,Lin J,et al.Ozone initiated inactivation of Escherichia coli and Staphylococcus aureus in water:Influence of selected organic solvents prevalent in wastewaters.Chemosphere,2018;206,43‐50.
18.Roy D,Wong PK,Engelbrecht RS,et al.Mechanism of enteroviral inactivation by ozone.Appl Environ Microbiol,1981;41,718‐23.
19.Kitamura N.[Genome structure of poliovirus(author's transl)].Seikagaku,1982;54,151‐6.
20.Simonet J,Gantzer C.Inactivation of poliovirus 1 and F‐specific RNA phages and degradation of their genomes by UVirradiation at 254 nanometers.Appl Environ Microbiol,2006;72,7671‐7.
21.Bhattacharya SS,Kulka M,Lampel KA,et al.Use of reverse transcription and PCR to discriminate between infectious and non‐infectious hepatitis A virus.J Virol Methods,2004;116,181‐7.
22.Jin M,Zhao ZG,Wang XW,et al.The 40‐80?nt region in the5'‐NCR of genome is a critical target for inactivating poliovirus by chlorine dioxide.J Med Virol,2012;84,526‐35.
23.Zaidi SS,Asghar H,Sharif S,et al.Poliovirus Laboratory Based Surveillance:An Overview.Methods Mol Biol,2016;1387,11‐8.
24.Katzenelson E,Biedermann N.Disinfection of viruses in sewage by ozone.Water Res,1976;10,629‐31.
25.Chatterjee NK,Kaehler M,Deibel R.Detection of enteroviruses using subgenomic probes of Coxsackie virus B4 by hybridization.Diagn Microbiol Infect Dis,1988;11,129‐36.
26.Cello J,Paul AV,Wimmer E.Chemical synthesis of poliovirus cDNA:generation of infectious virus in the absence of natural template.Science,2002;297,1016‐8.
27.Nuanualsuwan S,Cliver DO.Infectivity of RNA from inactivated poliovirus.Appl Environ Microbiol,2003;69,1629‐32.
28.Nomoto A,Omata T,Toyoda H,et al.Complete nucleotide sequence of the attenuated poliovirus Sabin 1 strain genome.Proc Natl Acad Sci USA,1982;79,5793‐7.
29.Camel V,Bermond A.The use of ozone and associated oxidation processes in drinking water treatment.Water Res,1998;32,3208‐22.
30.Moore NJ,Margolin AB.Efficacy of Nucleic Acid Probes for Detection of Poliovirus in Water Disinfected by Chlorine,Chlorine Dioxide,Ozone,and UV Radiation.Appl Environ Microbiol,1994;60,4189‐91.
31.Simonet J,Gantzer C.Degradation of the Poliovirus 1 genome by chlorine dioxide.J Appl Microbiol,2006;100,862‐70.
32.Li JW,Xin ZT,Wang XW,et al.Mechanisms of inactivation of hepatitis A virus in water by chlorine dioxide.Water Res,2004;38,1514‐9.
33.Souii A,Ben MM,Gharbi J.Role of RNA structure motifs in IRES‐dependent translation initiation of the coxsackievirus B3:new insights for developing live‐attenuated strains for vaccines and gene therapy.Mol Biotechnol,2013;55,179‐202.
34.Moustafa IM,Gohara DW,Uchida A,et al.Conformational Ensemble of the Poliovirus 3CD Precursor Observed by MDSimulations and Confirmed by SAXS:A Strategy to Expand the Viral Proteome.Viruses,2015;7,5962‐86.
35.Harris KS,Xiang W,Alexander L,et al.Interaction of poliovirus polypeptide 3CDpro with the 5'and 3'termini of the poliovirus genome.Identification of viral and cellular cofactors needed for efficient binding.J Biol Chem,1994;269,27004‐14.
36.Ogram SA,Spear A,Sharma N,et al.The 5'CL‐PCBP RNPcomplex,3'poly(A)tail and 2A(pro)are required for optimal translation of poliovirus RNA.Virology,2010;397,14‐22.
37.Schmid M,Wimmer E.IRES‐controlled protein synthesis and genome replication of poliovirus.Arch Virol Suppl,1994;9,279‐89.
38.Fitzgerald KD,Semler BL.Re‐localization of cellular protein SRp20 during poliovirus infection:bridging a viral IRES to the host cell translation apparatus.PLo S Pathog,2011;7,e1002127.
39.Muslin C,Joffret ML,Pelletier I,et al.Evolution and Emergence of Enteroviruses through Intra‐and Inter‐species Recombination:Plasticity and Phenotypic Impact of Modular Genetic Exchanges in the 5'Untranslated Region.PLoS Pathog,2015;11,e1005266.
40.Viktorova EG,Khattar S,Samal S,et al.Poliovirus Replicon RNAGeneration,Transfection,Packaging,and Quantitation of Replication.Curr Protoc Microbiol,2018;48,15H.4.1‐15H.4.15.
41.Ma JF,Straub TM,Pepper IL,et al.Cell culture and PCRdetermination of poliovirus inactivation by disinfectants.Appl Environ Microbiol,1994;60,4203‐6.
42.Maier,Tougianidou,Wiedenmann,et al.Detection of poliovirus by cell culture and by PCR after UV disinfection.Water Sci Technoly,1995;31,141‐5.
43.Lim MY,Kim JM,Ko G.Disinfection kinetics of murine norovirus using chlorine and chlorine dioxide.Water Res,2010;44,3243‐51.