The Experts below are selected from a list of 159 Experts worldwide ranked by ideXlab platform
Juan E Echevarria - One of the best experts on this subject based on the ideXlab platform.
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rapid molecular epidemiologic studies of Human Parainfluenza Viruses based on direct sequencing of amplified dna from a multiplex rt pcr assay
Journal of Virological Methods, 2000Co-Authors: Juan E Echevarria, Dean D Erdman, Cody H Meissner, Larry J AndersonAbstract:Sequencing studies of limited regions of the Human Parainfluenza Viruses (HPIVs) genomes have helped describe patterns of virus circulation and characterize institutional outbreaks of HPIVs-associated respiratory illness. In this study, we sequenced reverse transcription polymerase chain reaction (RT-PCR)-amplified HPIVs RNA obtained from a multiplex RT-PCR assay described previously for simultaneous detection of HPIV-1, 2 and 3. Differences in the nucleotide sequences of limited regions of the HN gene allowed us to distinguish temporally and geographically diverse HPIV isolates (43 HPIV-1, 7 HPIV-2, 12 HPIV-3 isolates from this and previously published studies). In addition, an outbreak of HPIV-3-associated illness among infants on a pediatric ward was investigated by comparing sequences of three ward isolates with three matched community controls. Sequences of all ward isolates were identical and differed from those of the community controls, suggesting a single introduction and nosocomial transmission of the virus. Combining multiplex reverse transcription polymerase chain reaction (RT-PCR) assays with direct sequencing of the PCR products can provide an integrated system for rapid diagnosis and characterization of HPIVs.
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detection and identification of Human Parainfluenza Viruses 1 2 3 and 4 in clinical samples of pediatric patients by multiplex reverse transcription pcr
Journal of Clinical Microbiology, 2000Co-Authors: Jose C Aguilar, Dean D Erdman, Maria P Perezbrena, Maria L Garcia, Nieves Cruz, Juan E EchevarriaAbstract:We describe a multiplex reverse transcription-PCR (m-RT-PCR) assay that is able to detect and differentiate all known Human Parainfluenza Viruses (HPIVs). Serial dilution experiments with reference strains that compared cell culture isolation and m-RT-PCR showed sensitivities ranging from 0.0004 50% tissue culture infective dose (TCID50) for HPIV type 4B (HPIV-4B) to 32 TCID50s for HPIV-3. As few as 10 plasmids containing HPIV PCR products could be detected in all cases. When 201 nasopharyngeal aspirate specimens from pediatric patients hospitalized for lower respiratory illness were tested, m-RT-PCR assay detected 64 HPIVs (24 HPIV-3, 23 HPIV-1, 10 HPIV-4, and 7 HPIV-2), while only 42 of them (21 HPIV-1, 14 HPIV-3, 6 HPIV-2, and 1 HPIV-4 isolates) grew in cell culture. Our m-RT-PCR assay was more sensitive than either cell culture isolation or indirect immunofluorescence with monoclonal antibodies for the detection of HPIV infections. Also, HPIV-4 was more frequently detected than HPIV-2 in this study, suggesting that it may have been underestimated as a lower respiratory tract pathogen because of the insensitivity of cell culture.
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simultaneous detection and identification of Human Parainfluenza Viruses 1 2 and 3 from clinical samples by multiplex pcr
Journal of Clinical Microbiology, 1998Co-Authors: Juan E Echevarria, Dean D Erdman, Ella M Swierkosz, Brian P Holloway, Larry J AndersonAbstract:Reverse transcription (RT)-PCR assays have been widely described for use in the diagnosis of Human Parainfluenza Viruses (HPIVs) and other respiratory virus pathogens. However, these assays are mostly monospecific, requiring separate amplifications for each HPIV type. In the present work, we describe multiplex RT-PCR assays that detect and differentiate HPIV serotypes 1, 2, and 3 in a combined reaction. Specifically, a mixture of three pairs of primers to conserved regions of the hemagglutinin-neuraminidase gene of each HPIV serotype was used for primary amplification, yielding amplicons with similar sizes. For typing, a second amplification was performed with a mixture of nested primers, yielding amplicons with sizes easily differentiated by agarose gel electrophoresis. A modified single-amplification RT-PCR assay with fluorescence-labeled nested primers, followed by analysis of the labeled products on an automated sequencing gel, was also evaluated. Fifteen temporally and geographically diverse HPIV isolates from the Centers for Disease Control and Prevention archives and 26 of 30 (87%) previously positive nasopharyngeal specimens (8 of 10 positive for HPIV serotype 1 [HPIV1], 9 of 10 positive for HPIV2, and 9 of 10 positive for HPIV3) were positive and were correctly typed by both assays. Negative results were obtained with naso- or oropharyngeal specimens and/or culture isolates of 33 unrelated respiratory tract pathogens, including HPIV4, enterovirus, rhinovirus, respiratory syncytial virus, adenovirus, influenza virus, and Streptococcus pneumoniae. Our multiplex RT-PCR assays provide sensitive, specific, and simplified tools for the rapid diagnosis of HPIV infections.
Anne Moscona - One of the best experts on this subject based on the ideXlab platform.
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A Recombinant Sialidase Fusion Protein Effectively Inhibits Human Parainfluenza Viral Infection in Vitro and in Vivo
The Journal of Infectious Diseases, 2010Co-Authors: Anne Moscona, Samantha G. Palmer, Lori Aschenbrenner, Gallen B. Triana-baltzer, Qi-xiang Li, David F. Wurtman, Stefan Niewiesk, Matteo Porotto, Fang FangAbstract:Background The first step in infection by Human Parainfluenza Viruses (HPIVs) is binding to the surface of respiratory epithelial cells via interaction between viral receptor-binding molecules and sialic acid-containing receptors. DAS181, a recombinant sialidase protein containing the catalytic domain of A. viscosus sialidase, removes cell surface sialic acid, and we proposed that it would inhibit HPIV infection.
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virus receptor interactions of Human Parainfluenza Viruses types 1 2 and 3
Microbial Pathogenesis, 1999Co-Authors: Collette Ahtye, Kety Huberman, Elizabeth Carlin, Stephanie Schwartz, Anne MosconaAbstract:Abstract Human Parainfluenza Viruses types 1, 2 and 3 (HPF 1, 2 and 3) are important pathogens in children. While these Viruses share common structures and replication strategies, they target different parts of the respiratory tract; the most common outcomes of infection with HPF3 are bronchiolitis and pneumonia, while HPF 1 and 2 are associated with croup. While the HPF3 fusion protein (F) is critical for membrane fusion, our previous work revealed that the receptor binding hemagglutinin–neuraminidase (HN) is also essential to the fusion process; interaction between HN and its sialic acid-containing receptor on cell surfaces is required for HPF3 mediated cell fusion. Using our understanding of HPF3 HN's functions in the cell-binding and viral entry process, we are investigating the ways in which these processes differ in HPF 1 and 2, in part by manipulating receptor availability. Three experimental treatments were used to compare the HN–receptor interaction of HPF 1, 2 and 3: infection at high multiplicity of infection (m.o.i.); bacterial neuraminidase treatment of cells infected at low m.o.i.; and viral neuraminidase treatment of cells infected at low m.o.i. (using Newcastle disease virus [NDV] neuraminidase or UV irradiated HPF3 as sources of neuraminidase). In cells infected with HPF3, we have shown that infection with high m.o.i. blocks fusion, by removing sialic acid receptors for the viral HN. However, in cells infected with HPF 1 and 2, infection with high m.o.i. did not block fusion; the fusion increases with increasing m.o.i.. In cells infected with HPF 1 and 2, neither bacterial nor NDV neuraminidase blocked cell fusion, using amounts of neuraminidase that completely block fusion of HPF3 infected cells. However, when inactivated HPF3 was used as a source of viral neuraminidase, the treatment inhibited fusion of cells infected with HPF 1 and 2 as well as 3. The differences found between these Viruses in terms of their interaction with the cell, ability to modulate cell–cell fusion and reponse to exogenous neuraminidases of various specificities, may reflect salient differences in biological properties of the three Viruses.
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comparison of the virus receptor interactions of Human Parainfluenza Viruses types 1 2 and 3 995
Pediatric Research, 1996Co-Authors: Elizabeth Carlin, Kety Huberman, Collette Ahtye, Stephanie Schwartz, Anne MosconaAbstract:COMPARISON OF THE VIRUS-RECEPTOR INTERACTIONS OF Human Parainfluenza Viruses TYPES 1, 2 and 3. † 995
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COMPARISON OF THE VIRUS-RECEPTOR INTERACTIONS OF Human Parainfluenza Viruses TYPES 1, 2 and 3. † 995
Pediatric Research, 1996Co-Authors: Elizabeth Carlin, Kety Huberman, Stephanie Schwartz, Anne MosconaAbstract:COMPARISON OF THE VIRUS-RECEPTOR INTERACTIONS OF Human Parainfluenza Viruses TYPES 1, 2 and 3. † 995
Dean D Erdman - One of the best experts on this subject based on the ideXlab platform.
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Hospitalization due to Human Parainfluenza virus–associated lower respiratory tract illness in rural Thailand
Influenza and Other Respiratory Viruses, 2012Co-Authors: Oliver Morgan, Malinee Chittaganpitch, Birgit Clague, Somrak Chantra, Wichai Sanasuttipun, Prabda Prapasiri, Sathapana Naorat, Yongjua Laosirithavorn, Teresa C. T. Peret, Dean D ErdmanAbstract:Background Human Parainfluenza Viruses (HPIVs) are an important cause of acute respiratory illness in young children but little is known about their epidemiology in the tropics. Methods From 2003–2007, we conducted surveillance for hospitalized respiratory illness in rural Thailand. We performed reverse-transcriptase polymerase chain reaction on nasopharyngeal specimens and enzyme immunoassay on paired sera Results Of 10,097 patients enrolled, 573 (5%) of all ages and 370 (9%) of children
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rapid molecular epidemiologic studies of Human Parainfluenza Viruses based on direct sequencing of amplified dna from a multiplex rt pcr assay
Journal of Virological Methods, 2000Co-Authors: Juan E Echevarria, Dean D Erdman, Cody H Meissner, Larry J AndersonAbstract:Sequencing studies of limited regions of the Human Parainfluenza Viruses (HPIVs) genomes have helped describe patterns of virus circulation and characterize institutional outbreaks of HPIVs-associated respiratory illness. In this study, we sequenced reverse transcription polymerase chain reaction (RT-PCR)-amplified HPIVs RNA obtained from a multiplex RT-PCR assay described previously for simultaneous detection of HPIV-1, 2 and 3. Differences in the nucleotide sequences of limited regions of the HN gene allowed us to distinguish temporally and geographically diverse HPIV isolates (43 HPIV-1, 7 HPIV-2, 12 HPIV-3 isolates from this and previously published studies). In addition, an outbreak of HPIV-3-associated illness among infants on a pediatric ward was investigated by comparing sequences of three ward isolates with three matched community controls. Sequences of all ward isolates were identical and differed from those of the community controls, suggesting a single introduction and nosocomial transmission of the virus. Combining multiplex reverse transcription polymerase chain reaction (RT-PCR) assays with direct sequencing of the PCR products can provide an integrated system for rapid diagnosis and characterization of HPIVs.
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detection and identification of Human Parainfluenza Viruses 1 2 3 and 4 in clinical samples of pediatric patients by multiplex reverse transcription pcr
Journal of Clinical Microbiology, 2000Co-Authors: Jose C Aguilar, Dean D Erdman, Maria P Perezbrena, Maria L Garcia, Nieves Cruz, Juan E EchevarriaAbstract:We describe a multiplex reverse transcription-PCR (m-RT-PCR) assay that is able to detect and differentiate all known Human Parainfluenza Viruses (HPIVs). Serial dilution experiments with reference strains that compared cell culture isolation and m-RT-PCR showed sensitivities ranging from 0.0004 50% tissue culture infective dose (TCID50) for HPIV type 4B (HPIV-4B) to 32 TCID50s for HPIV-3. As few as 10 plasmids containing HPIV PCR products could be detected in all cases. When 201 nasopharyngeal aspirate specimens from pediatric patients hospitalized for lower respiratory illness were tested, m-RT-PCR assay detected 64 HPIVs (24 HPIV-3, 23 HPIV-1, 10 HPIV-4, and 7 HPIV-2), while only 42 of them (21 HPIV-1, 14 HPIV-3, 6 HPIV-2, and 1 HPIV-4 isolates) grew in cell culture. Our m-RT-PCR assay was more sensitive than either cell culture isolation or indirect immunofluorescence with monoclonal antibodies for the detection of HPIV infections. Also, HPIV-4 was more frequently detected than HPIV-2 in this study, suggesting that it may have been underestimated as a lower respiratory tract pathogen because of the insensitivity of cell culture.
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simultaneous detection and identification of Human Parainfluenza Viruses 1 2 and 3 from clinical samples by multiplex pcr
Journal of Clinical Microbiology, 1998Co-Authors: Juan E Echevarria, Dean D Erdman, Ella M Swierkosz, Brian P Holloway, Larry J AndersonAbstract:Reverse transcription (RT)-PCR assays have been widely described for use in the diagnosis of Human Parainfluenza Viruses (HPIVs) and other respiratory virus pathogens. However, these assays are mostly monospecific, requiring separate amplifications for each HPIV type. In the present work, we describe multiplex RT-PCR assays that detect and differentiate HPIV serotypes 1, 2, and 3 in a combined reaction. Specifically, a mixture of three pairs of primers to conserved regions of the hemagglutinin-neuraminidase gene of each HPIV serotype was used for primary amplification, yielding amplicons with similar sizes. For typing, a second amplification was performed with a mixture of nested primers, yielding amplicons with sizes easily differentiated by agarose gel electrophoresis. A modified single-amplification RT-PCR assay with fluorescence-labeled nested primers, followed by analysis of the labeled products on an automated sequencing gel, was also evaluated. Fifteen temporally and geographically diverse HPIV isolates from the Centers for Disease Control and Prevention archives and 26 of 30 (87%) previously positive nasopharyngeal specimens (8 of 10 positive for HPIV serotype 1 [HPIV1], 9 of 10 positive for HPIV2, and 9 of 10 positive for HPIV3) were positive and were correctly typed by both assays. Negative results were obtained with naso- or oropharyngeal specimens and/or culture isolates of 33 unrelated respiratory tract pathogens, including HPIV4, enterovirus, rhinovirus, respiratory syncytial virus, adenovirus, influenza virus, and Streptococcus pneumoniae. Our multiplex RT-PCR assays provide sensitive, specific, and simplified tools for the rapid diagnosis of HPIV infections.
Larry J Anderson - One of the best experts on this subject based on the ideXlab platform.
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rapid molecular epidemiologic studies of Human Parainfluenza Viruses based on direct sequencing of amplified dna from a multiplex rt pcr assay
Journal of Virological Methods, 2000Co-Authors: Juan E Echevarria, Dean D Erdman, Cody H Meissner, Larry J AndersonAbstract:Sequencing studies of limited regions of the Human Parainfluenza Viruses (HPIVs) genomes have helped describe patterns of virus circulation and characterize institutional outbreaks of HPIVs-associated respiratory illness. In this study, we sequenced reverse transcription polymerase chain reaction (RT-PCR)-amplified HPIVs RNA obtained from a multiplex RT-PCR assay described previously for simultaneous detection of HPIV-1, 2 and 3. Differences in the nucleotide sequences of limited regions of the HN gene allowed us to distinguish temporally and geographically diverse HPIV isolates (43 HPIV-1, 7 HPIV-2, 12 HPIV-3 isolates from this and previously published studies). In addition, an outbreak of HPIV-3-associated illness among infants on a pediatric ward was investigated by comparing sequences of three ward isolates with three matched community controls. Sequences of all ward isolates were identical and differed from those of the community controls, suggesting a single introduction and nosocomial transmission of the virus. Combining multiplex reverse transcription polymerase chain reaction (RT-PCR) assays with direct sequencing of the PCR products can provide an integrated system for rapid diagnosis and characterization of HPIVs.
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simultaneous detection and identification of Human Parainfluenza Viruses 1 2 and 3 from clinical samples by multiplex pcr
Journal of Clinical Microbiology, 1998Co-Authors: Juan E Echevarria, Dean D Erdman, Ella M Swierkosz, Brian P Holloway, Larry J AndersonAbstract:Reverse transcription (RT)-PCR assays have been widely described for use in the diagnosis of Human Parainfluenza Viruses (HPIVs) and other respiratory virus pathogens. However, these assays are mostly monospecific, requiring separate amplifications for each HPIV type. In the present work, we describe multiplex RT-PCR assays that detect and differentiate HPIV serotypes 1, 2, and 3 in a combined reaction. Specifically, a mixture of three pairs of primers to conserved regions of the hemagglutinin-neuraminidase gene of each HPIV serotype was used for primary amplification, yielding amplicons with similar sizes. For typing, a second amplification was performed with a mixture of nested primers, yielding amplicons with sizes easily differentiated by agarose gel electrophoresis. A modified single-amplification RT-PCR assay with fluorescence-labeled nested primers, followed by analysis of the labeled products on an automated sequencing gel, was also evaluated. Fifteen temporally and geographically diverse HPIV isolates from the Centers for Disease Control and Prevention archives and 26 of 30 (87%) previously positive nasopharyngeal specimens (8 of 10 positive for HPIV serotype 1 [HPIV1], 9 of 10 positive for HPIV2, and 9 of 10 positive for HPIV3) were positive and were correctly typed by both assays. Negative results were obtained with naso- or oropharyngeal specimens and/or culture isolates of 33 unrelated respiratory tract pathogens, including HPIV4, enterovirus, rhinovirus, respiratory syncytial virus, adenovirus, influenza virus, and Streptococcus pneumoniae. Our multiplex RT-PCR assays provide sensitive, specific, and simplified tools for the rapid diagnosis of HPIV infections.
Mark Von Itzstein - One of the best experts on this subject based on the ideXlab platform.
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The impact of the butterfly effect on Human Parainfluenza virus haemagglutinin-neuraminidase inhibitor design
Scientific Reports, 2017Co-Authors: Larissa Dirr, Ibrahim M. El-deeb, Leonard M. G. Chavas, Patrice Guillon, Mark Von ItzsteinAbstract:Human Parainfluenza Viruses represent a leading cause of lower respiratory tract disease in children, with currently no available approved drug or vaccine. The viral surface glycoprotein haemagglutinin-neuraminidase (HN) represents an ideal antiviral target. Herein, we describe the first structure-based study on the rearrangement of key active site amino acid residues by an induced opening of the 216-loop, through the accommodation of appropriately functionalised neuraminic acid-based inhibitors. We discovered that the rearrangement is influenced by the degree of loop opening and is controlled by the neuraminic acid’s C-4 substituent’s size (large or small). In this study, we found that these rearrangements induce a butterfly effect of paramount importance in HN inhibitor design and define criteria for the ideal substituent size in two different categories of HN inhibitors and provide novel structural insight into the druggable viral HN protein.
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Structure-guided discovery of potent and dual-acting Human Parainfluenza virus haemagglutinin–neuraminidase inhibitors
Nature Communications, 2014Co-Authors: Patrice Guillon, Larissa Dirr, Ibrahim M. El-deeb, Moritz Winger, Benjamin Bailly, Thomas Haselhorst, Jeffrey Clifford Dyason, Mark Von ItzsteinAbstract:Human Parainfluenza Viruses (hPIVs) cause common respiratory diseases in children. Here the authors rationally design small molecules targeting the hPIV haemagglutinin–neuraminidase protein, and show that the compounds inhibit viral entry and exit from cultured cells.
