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Sagar M Goyal - One of the best experts on this subject based on the ideXlab platform.
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Phylogenetic analysis, genomic diversity and classification of M class gene segments of turkey Reoviruses.
Veterinary microbiology, 2015Co-Authors: Sunil K Mor, Douglas Marthaler, Harsha Verma, Tamer A Sharafeldin, Naresh Jindal, Robert E Porter, Sagar M GoyalAbstract:From 2011 to 2014, 13 turkey arthritis Reoviruses (TARVs) were isolated from cases of swollen hock joints in 2-18-week-old turkeys. In addition, two isolates from similar cases of turkey arthritis were received from another laboratory. Eight turkey enteric Reoviruses (TERVs) isolated from fecal samples of turkeys were also used for comparison. The aims of this study were to characterize turkey reovirus (TRV) based on complete M class genome segments and to determine genetic diversity within TARVs in comparison to TERVs and chicken Reoviruses (CRVs). Nucleotide (nt) cut off values of 84%, 83% and 85% for the M1, M2 and M3 gene segments were proposed and used for genotype classification, generating 5, 7, and 3 genotypes, respectively. Using these nt cut off values, we propose M class genotype constellations (GCs) for avian Reoviruses. Of the seven GCs, GC1 and GC3 were shared between the TARVs and TERVs, indicating possible reassortment between turkey and chicken Reoviruses. The TARVs and TERVs were divided into three GCs, and GC2 was unique to TARVs and TERVs. The proposed new GC approach should be useful in identifying reassortant viruses, which may ultimately be used in the design of a universal vaccine against both chicken and turkey Reoviruses.
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one step real time reverse transcription pcr for the detection of turkey Reoviruses
Avian Diseases, 2014Co-Authors: Sunil K Mor, Harsha Verma, Tamer A Sharafeldin, Robert E Porter, Aschalew Z. Bekele, Sagar M GoyalAbstract:SUMMARY During late 2010 and early 2011, an unusual problem of lameness and swollen hock joints in commercial turkeys was reported in the upper Midwest, which continues to this day. The disease caused substantial economic losses to turkey producers. Reovirus was isolated from tendons and joint fluids of lame turkeys submitted to the Minnesota Veterinary Diagnostic Laboratory. This study was undertaken to develop a TaqMan real-time reverse transcription-PCR (rRT-PCR) assay for the early detection of turkey Reoviruses (both enteric and lameness strains). A primer probe set was designed from the conserved region of the S4 segment of the turkey reovirus genome. The newly developed rRT-PCR was specific for the detection of turkey Reoviruses. The detection limit of this assay was 10 genome copies per reaction. For the TARV-MN4 strain of turkey arthritis reovirus, one 50% tissue culture infectious dose was equivalent to 11.6 ± 0.2 genome copies. The highest coefficient of variation for intraexperimental and inte...
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One-Step Real-Time Reverse Transcription–PCR for the Detection of Turkey Reoviruses
Avian diseases, 2014Co-Authors: Sunil K Mor, Harsha Verma, Tamer A Sharafeldin, Robert E Porter, Aschalew Z. Bekele, Sagar M GoyalAbstract:SUMMARY During late 2010 and early 2011, an unusual problem of lameness and swollen hock joints in commercial turkeys was reported in the upper Midwest, which continues to this day. The disease caused substantial economic losses to turkey producers. Reovirus was isolated from tendons and joint fluids of lame turkeys submitted to the Minnesota Veterinary Diagnostic Laboratory. This study was undertaken to develop a TaqMan real-time reverse transcription-PCR (rRT-PCR) assay for the early detection of turkey Reoviruses (both enteric and lameness strains). A primer probe set was designed from the conserved region of the S4 segment of the turkey reovirus genome. The newly developed rRT-PCR was specific for the detection of turkey Reoviruses. The detection limit of this assay was 10 genome copies per reaction. For the TARV-MN4 strain of turkey arthritis reovirus, one 50% tissue culture infectious dose was equivalent to 11.6 ± 0.2 genome copies. The highest coefficient of variation for intraexperimental and inte...
Bernard N. Fields - One of the best experts on this subject based on the ideXlab platform.
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A pathway for entry of Reoviruses into the host through M cells of the respiratory tract.
The Journal of experimental medicine, 1994Co-Authors: M J Morin, A Warner, Bernard N. FieldsAbstract:Many microorganisms gain access to the systemic circulation after entering the respiratory tract. The precise pathways used to cross the mucosal barriers of the lungs have not been clearly described. We have used the mammalian Reoviruses in order to determine the pathway that a systemic virus uses to penetrate the mucosal barrier and enter the systemic circulation after entering the airways of the lungs. Reoviruses enter through pulmonary M cells, which overlie bronchus-associated lymphoid tissue, and subsequently spread to regional lymph nodes. Thus, the pathway through M cells represents a strategy by which viruses and probably other microorganisms can penetrate the mucosal surface of the respiratory tract and thereby enter the systemic circulation.
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A Carboxy-Terminal Fragment ofProtein ,ul/,uC IsPresent inInfectious Subvirion Particles ofMammalian Reoviruses andIsProposed ToHavea RoleinPenetration
1992Co-Authors: Max L. Nibert, Bernard N. FieldsAbstract:Penetration ofa cell membrane asan earlyeventininfection ofcells bymammalian Reoviruses appearsto require aparticular typeofviral particle, theinfectious subvirion particle (ISVP), whichisgenerated froman intact virion byproteolytic cleavage oftheoutercapsid proteins o3andul1/ulC. Characterizations ofthe structural components andproperties ofISVPs arethusrelevant toattempts tounderstand themechanism of penetration byReoviruses. Inthis study, anovel, -13-kDa carboxy-terminal fragment (given thename +)was found tobegenerated fromprotein pl/plC during invitro treatments ofvirions withtrypsin orchymotrypsin toyield ISVPs.Withtrypsin treatment, boththecarboxy-terminal fragment + andtheamino-terminal fragment p18/8 wereshowntobegenerated andtoremain attached toISVPsinstoichiometric quantities. Sites ofprotease cleavage were identified inthededucedaminoacidsequenceof,ulbydetermining the amino-terminal sequencesof+ proteins: trypsin cleaves between arginine 584andisoleucine 585,and chymotrypsin cleaves between tyrosine 581andglycine 582.Findings inthis study indicate that sequencesin the+ portion of,l1/,ulC may participate intheunique functions attributed toISVPs.Notably, the8-+ cleavage junction was predicted tobeflanked bya pairoflongamphipathic at-helices. Theseamphipathic a-helices, together withthemyristoyl groupattheextreme aminoterminus of,uV/lIN, areproposed tointeract directly withthelipid bilayer ofa cell membraneduring penetration bymammalian Reoviruses. Themechanisms bywhichdifferent nonenveloped viruses penetrate cell membranes toenterthecytoplasm early in infection ofcells arepoorly understood. Itisimportant to understand these mechanisms since thestepsrequired to makeviral particles competent forpenetration andthesteps inpenetration itself arelikely toserve asdeterminants ofcell ortissue tropism andinjury andthustohaverelevance to studies ofviral pathogenesis (33, 50). We haveundertaken experiments that attempt todefine this mechanism forone particular groupofnonenveloped viruses, themammalian Reoviruses.
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A carboxy-terminal fragment of protein mu 1/mu 1C is present in infectious subvirion particles of mammalian Reoviruses and is proposed to have a role in penetration.
Journal of virology, 1992Co-Authors: Max L. Nibert, Bernard N. FieldsAbstract:Penetration of a cell membrane as an early event in infection of cells by mammalian Reoviruses appears to require a particular type of viral particle, the infectious subvirion particle (ISVP), which is generated from an intact virion by proteolytic cleavage of the outer capsid proteins sigma 3 and mu 1/mu 1C. Characterizations of the structural components and properties of ISVPs are thus relevant to attempts to understand the mechanism of penetration by Reoviruses. In this study, a novel, approximately 13-kDa carboxy-terminal fragment (given the name phi) was found to be generated from protein mu 1/mu 1C during in vitro treatments of virions with trypsin or chymotrypsin to yield ISVPs. With trypsin treatment, both the carboxy-terminal fragment phi and the amino-terminal fragment mu 1 delta/delta were shown to be generated and to remain attached to ISVPs in stoichiometric quantities. Sites of protease cleavage were identified in the deduced amino acid sequence of mu 1 by determining the amino-terminal sequences of phi proteins: trypsin cleaves between arginine 584 and isoleucine 585, and chymotrypsin cleaves between tyrosine 581 and glycine 582. Findings in this study indicate that sequences in the phi portion of mu 1/mu 1C may participate in the unique functions attributed to ISVPs. Notably, the delta-phi cleavage junction was predicted to be flanked by a pair of long amphipathic alpha-helices. These amphipathic alpha-helices, together with the myristoyl group at the extreme amino terminus of mu 1/mu 1N, are proposed to interact directly with the lipid bilayer of a cell membrane during penetration by mammalian Reoviruses.
Sunil K Mor - One of the best experts on this subject based on the ideXlab platform.
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Phylogenetic analysis, genomic diversity and classification of M class gene segments of turkey Reoviruses.
Veterinary microbiology, 2015Co-Authors: Sunil K Mor, Douglas Marthaler, Harsha Verma, Tamer A Sharafeldin, Naresh Jindal, Robert E Porter, Sagar M GoyalAbstract:From 2011 to 2014, 13 turkey arthritis Reoviruses (TARVs) were isolated from cases of swollen hock joints in 2-18-week-old turkeys. In addition, two isolates from similar cases of turkey arthritis were received from another laboratory. Eight turkey enteric Reoviruses (TERVs) isolated from fecal samples of turkeys were also used for comparison. The aims of this study were to characterize turkey reovirus (TRV) based on complete M class genome segments and to determine genetic diversity within TARVs in comparison to TERVs and chicken Reoviruses (CRVs). Nucleotide (nt) cut off values of 84%, 83% and 85% for the M1, M2 and M3 gene segments were proposed and used for genotype classification, generating 5, 7, and 3 genotypes, respectively. Using these nt cut off values, we propose M class genotype constellations (GCs) for avian Reoviruses. Of the seven GCs, GC1 and GC3 were shared between the TARVs and TERVs, indicating possible reassortment between turkey and chicken Reoviruses. The TARVs and TERVs were divided into three GCs, and GC2 was unique to TARVs and TERVs. The proposed new GC approach should be useful in identifying reassortant viruses, which may ultimately be used in the design of a universal vaccine against both chicken and turkey Reoviruses.
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one step real time reverse transcription pcr for the detection of turkey Reoviruses
Avian Diseases, 2014Co-Authors: Sunil K Mor, Harsha Verma, Tamer A Sharafeldin, Robert E Porter, Aschalew Z. Bekele, Sagar M GoyalAbstract:SUMMARY During late 2010 and early 2011, an unusual problem of lameness and swollen hock joints in commercial turkeys was reported in the upper Midwest, which continues to this day. The disease caused substantial economic losses to turkey producers. Reovirus was isolated from tendons and joint fluids of lame turkeys submitted to the Minnesota Veterinary Diagnostic Laboratory. This study was undertaken to develop a TaqMan real-time reverse transcription-PCR (rRT-PCR) assay for the early detection of turkey Reoviruses (both enteric and lameness strains). A primer probe set was designed from the conserved region of the S4 segment of the turkey reovirus genome. The newly developed rRT-PCR was specific for the detection of turkey Reoviruses. The detection limit of this assay was 10 genome copies per reaction. For the TARV-MN4 strain of turkey arthritis reovirus, one 50% tissue culture infectious dose was equivalent to 11.6 ± 0.2 genome copies. The highest coefficient of variation for intraexperimental and inte...
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One-Step Real-Time Reverse Transcription–PCR for the Detection of Turkey Reoviruses
Avian diseases, 2014Co-Authors: Sunil K Mor, Harsha Verma, Tamer A Sharafeldin, Robert E Porter, Aschalew Z. Bekele, Sagar M GoyalAbstract:SUMMARY During late 2010 and early 2011, an unusual problem of lameness and swollen hock joints in commercial turkeys was reported in the upper Midwest, which continues to this day. The disease caused substantial economic losses to turkey producers. Reovirus was isolated from tendons and joint fluids of lame turkeys submitted to the Minnesota Veterinary Diagnostic Laboratory. This study was undertaken to develop a TaqMan real-time reverse transcription-PCR (rRT-PCR) assay for the early detection of turkey Reoviruses (both enteric and lameness strains). A primer probe set was designed from the conserved region of the S4 segment of the turkey reovirus genome. The newly developed rRT-PCR was specific for the detection of turkey Reoviruses. The detection limit of this assay was 10 genome copies per reaction. For the TARV-MN4 strain of turkey arthritis reovirus, one 50% tissue culture infectious dose was equivalent to 11.6 ± 0.2 genome copies. The highest coefficient of variation for intraexperimental and inte...
Robert E Porter - One of the best experts on this subject based on the ideXlab platform.
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Phylogenetic analysis, genomic diversity and classification of M class gene segments of turkey Reoviruses.
Veterinary microbiology, 2015Co-Authors: Sunil K Mor, Douglas Marthaler, Harsha Verma, Tamer A Sharafeldin, Naresh Jindal, Robert E Porter, Sagar M GoyalAbstract:From 2011 to 2014, 13 turkey arthritis Reoviruses (TARVs) were isolated from cases of swollen hock joints in 2-18-week-old turkeys. In addition, two isolates from similar cases of turkey arthritis were received from another laboratory. Eight turkey enteric Reoviruses (TERVs) isolated from fecal samples of turkeys were also used for comparison. The aims of this study were to characterize turkey reovirus (TRV) based on complete M class genome segments and to determine genetic diversity within TARVs in comparison to TERVs and chicken Reoviruses (CRVs). Nucleotide (nt) cut off values of 84%, 83% and 85% for the M1, M2 and M3 gene segments were proposed and used for genotype classification, generating 5, 7, and 3 genotypes, respectively. Using these nt cut off values, we propose M class genotype constellations (GCs) for avian Reoviruses. Of the seven GCs, GC1 and GC3 were shared between the TARVs and TERVs, indicating possible reassortment between turkey and chicken Reoviruses. The TARVs and TERVs were divided into three GCs, and GC2 was unique to TARVs and TERVs. The proposed new GC approach should be useful in identifying reassortant viruses, which may ultimately be used in the design of a universal vaccine against both chicken and turkey Reoviruses.
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one step real time reverse transcription pcr for the detection of turkey Reoviruses
Avian Diseases, 2014Co-Authors: Sunil K Mor, Harsha Verma, Tamer A Sharafeldin, Robert E Porter, Aschalew Z. Bekele, Sagar M GoyalAbstract:SUMMARY During late 2010 and early 2011, an unusual problem of lameness and swollen hock joints in commercial turkeys was reported in the upper Midwest, which continues to this day. The disease caused substantial economic losses to turkey producers. Reovirus was isolated from tendons and joint fluids of lame turkeys submitted to the Minnesota Veterinary Diagnostic Laboratory. This study was undertaken to develop a TaqMan real-time reverse transcription-PCR (rRT-PCR) assay for the early detection of turkey Reoviruses (both enteric and lameness strains). A primer probe set was designed from the conserved region of the S4 segment of the turkey reovirus genome. The newly developed rRT-PCR was specific for the detection of turkey Reoviruses. The detection limit of this assay was 10 genome copies per reaction. For the TARV-MN4 strain of turkey arthritis reovirus, one 50% tissue culture infectious dose was equivalent to 11.6 ± 0.2 genome copies. The highest coefficient of variation for intraexperimental and inte...
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One-Step Real-Time Reverse Transcription–PCR for the Detection of Turkey Reoviruses
Avian diseases, 2014Co-Authors: Sunil K Mor, Harsha Verma, Tamer A Sharafeldin, Robert E Porter, Aschalew Z. Bekele, Sagar M GoyalAbstract:SUMMARY During late 2010 and early 2011, an unusual problem of lameness and swollen hock joints in commercial turkeys was reported in the upper Midwest, which continues to this day. The disease caused substantial economic losses to turkey producers. Reovirus was isolated from tendons and joint fluids of lame turkeys submitted to the Minnesota Veterinary Diagnostic Laboratory. This study was undertaken to develop a TaqMan real-time reverse transcription-PCR (rRT-PCR) assay for the early detection of turkey Reoviruses (both enteric and lameness strains). A primer probe set was designed from the conserved region of the S4 segment of the turkey reovirus genome. The newly developed rRT-PCR was specific for the detection of turkey Reoviruses. The detection limit of this assay was 10 genome copies per reaction. For the TARV-MN4 strain of turkey arthritis reovirus, one 50% tissue culture infectious dose was equivalent to 11.6 ± 0.2 genome copies. The highest coefficient of variation for intraexperimental and inte...
Tamer A Sharafeldin - One of the best experts on this subject based on the ideXlab platform.
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Phylogenetic analysis, genomic diversity and classification of M class gene segments of turkey Reoviruses.
Veterinary microbiology, 2015Co-Authors: Sunil K Mor, Douglas Marthaler, Harsha Verma, Tamer A Sharafeldin, Naresh Jindal, Robert E Porter, Sagar M GoyalAbstract:From 2011 to 2014, 13 turkey arthritis Reoviruses (TARVs) were isolated from cases of swollen hock joints in 2-18-week-old turkeys. In addition, two isolates from similar cases of turkey arthritis were received from another laboratory. Eight turkey enteric Reoviruses (TERVs) isolated from fecal samples of turkeys were also used for comparison. The aims of this study were to characterize turkey reovirus (TRV) based on complete M class genome segments and to determine genetic diversity within TARVs in comparison to TERVs and chicken Reoviruses (CRVs). Nucleotide (nt) cut off values of 84%, 83% and 85% for the M1, M2 and M3 gene segments were proposed and used for genotype classification, generating 5, 7, and 3 genotypes, respectively. Using these nt cut off values, we propose M class genotype constellations (GCs) for avian Reoviruses. Of the seven GCs, GC1 and GC3 were shared between the TARVs and TERVs, indicating possible reassortment between turkey and chicken Reoviruses. The TARVs and TERVs were divided into three GCs, and GC2 was unique to TARVs and TERVs. The proposed new GC approach should be useful in identifying reassortant viruses, which may ultimately be used in the design of a universal vaccine against both chicken and turkey Reoviruses.
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one step real time reverse transcription pcr for the detection of turkey Reoviruses
Avian Diseases, 2014Co-Authors: Sunil K Mor, Harsha Verma, Tamer A Sharafeldin, Robert E Porter, Aschalew Z. Bekele, Sagar M GoyalAbstract:SUMMARY During late 2010 and early 2011, an unusual problem of lameness and swollen hock joints in commercial turkeys was reported in the upper Midwest, which continues to this day. The disease caused substantial economic losses to turkey producers. Reovirus was isolated from tendons and joint fluids of lame turkeys submitted to the Minnesota Veterinary Diagnostic Laboratory. This study was undertaken to develop a TaqMan real-time reverse transcription-PCR (rRT-PCR) assay for the early detection of turkey Reoviruses (both enteric and lameness strains). A primer probe set was designed from the conserved region of the S4 segment of the turkey reovirus genome. The newly developed rRT-PCR was specific for the detection of turkey Reoviruses. The detection limit of this assay was 10 genome copies per reaction. For the TARV-MN4 strain of turkey arthritis reovirus, one 50% tissue culture infectious dose was equivalent to 11.6 ± 0.2 genome copies. The highest coefficient of variation for intraexperimental and inte...
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One-Step Real-Time Reverse Transcription–PCR for the Detection of Turkey Reoviruses
Avian diseases, 2014Co-Authors: Sunil K Mor, Harsha Verma, Tamer A Sharafeldin, Robert E Porter, Aschalew Z. Bekele, Sagar M GoyalAbstract:SUMMARY During late 2010 and early 2011, an unusual problem of lameness and swollen hock joints in commercial turkeys was reported in the upper Midwest, which continues to this day. The disease caused substantial economic losses to turkey producers. Reovirus was isolated from tendons and joint fluids of lame turkeys submitted to the Minnesota Veterinary Diagnostic Laboratory. This study was undertaken to develop a TaqMan real-time reverse transcription-PCR (rRT-PCR) assay for the early detection of turkey Reoviruses (both enteric and lameness strains). A primer probe set was designed from the conserved region of the S4 segment of the turkey reovirus genome. The newly developed rRT-PCR was specific for the detection of turkey Reoviruses. The detection limit of this assay was 10 genome copies per reaction. For the TARV-MN4 strain of turkey arthritis reovirus, one 50% tissue culture infectious dose was equivalent to 11.6 ± 0.2 genome copies. The highest coefficient of variation for intraexperimental and inte...
