The Experts below are selected from a list of 120 Experts worldwide ranked by ideXlab platform

Duncan R. Smith - One of the best experts on this subject based on the ideXlab platform.

  • Amino acid alignment of flaviVirus NS5 proteins.
    2018
    Co-Authors: Chonticha Saisawang, Atichat Kuadkitkan, Prasert Auewarakul, Duncan R. Smith, Albert J. Ketterman
    Abstract:

    The sequences are nonstructural protein NS5 from Dengue Virus 2 NP_739590.2, Dengue Virus 1 NP_722465.1, Dengue Virus 3 YP_001531176.2, Dengue Virus 4 NP_740325.1, Zika Virus AMR39834.1, Japanese encephalitis Virus NP_775674.1, West Nile Virus YP_001527887.1, Tick-borne encephalitis Virus NP_775511.1, Yellow fever Virus NP_776009.1, Murray Valley encephalitis Virus NP_722539.1, Langat Virus NP_740302.1. Cysteines are highlighted in yellow with the positions of the 3 glutathionylated cysteines identified in this study shown in green highlight in the alignment. Blue bars illustrate the residues of the finger subdomains, the red bars show the residues of the palm domain and the purple bars show the residues of the thumb domain [30]. Conservation of amino acid sequence is shown by 100 percent conserved as white on black, between 80 to 100 percent conserved as white on dark grey, between 60 to 80 percent conserved as black on light grey, and less than 60 percent conserved as black on white.

  • Delayed antibody dependent enhancement of low passage Dengue Virus 4 isolates
    BMC Research Notes, 2015
    Co-Authors: Nitwara Wikan, Prasert Auewarakul, Sirikwan Libsittikul, Sutee Yoksan, Duncan R. Smith
    Abstract:

    Background The concept of antibody dependent enhancement (ADE) of Dengue Virus (DENV) infection is a cornerstone of our current understanding of Dengue pathogenesis, although some questions as to the mechanism remain, particularly in regards to the behavior of low and high passage Virus isolates. This study utilized two low passage DENV 4 isolates and a laboratory adapted DENV 4 isolate to investigate the potential of low passage isolates to undergo ADE. Results Little or no ADE of infection was observed on day 2 post infection with low passage isolates, while high enhancement of infection was seen with the laboratory adapted Virus. However, both of the low passage isolates showed high levels of infection (60–100 %) by day 5 post infection. Conclusions These results show that low passage DENV 4 Viruses undergo ADE mediated infection, but that the process is significantly temporally delayed as compared to laboratory adapted DENV 4.

  • Delayed antibody dependent enhancement of low passage Dengue Virus 4 isolates
    BMC research notes, 2015
    Co-Authors: Nitwara Wikan, Prasert Auewarakul, Sirikwan Libsittikul, Sutee Yoksan, Duncan R. Smith
    Abstract:

    Background The concept of antibody dependent enhancement (ADE) of Dengue Virus (DENV) infection is a cornerstone of our current understanding of Dengue pathogenesis, although some questions as to the mechanism remain, particularly in regards to the behavior of low and high passage Virus isolates. This study utilized two low passage DENV 4 isolates and a laboratory adapted DENV 4 isolate to investigate the potential of low passage isolates to undergo ADE.

Elizabeth Hunsperger - One of the best experts on this subject based on the ideXlab platform.

  • Development and characterization of serotype-specific monoclonal antibodies against the Dengue Virus-4 (DENV-4) non-structural protein (NS1)
    Virology Journal, 2018
    Co-Authors: Tesfaye Gelanew, Elizabeth Hunsperger
    Abstract:

    Background Dengue, caused by one of the four serologically distinct Dengue Viruses (DENV-1 to − 4), is a mosquito-borne disease of serious global health significance. Reliable and cost-effective diagnostic tests, along with effective vaccines and vector-control strategies, are highly required to reduce Dengue morbidity and mortality. Evaluation studies revealed that many commercially available NS1 antigen (Ag) tests have limited sensitivity to DENV-4 serotype compared to the other three serotypes. These studies indicated the need for development of new NS1 Ag detection test with improved sensitivity to DENV-4. An NS1 capture enzyme linked immunoassay (ELISA) specific to DENV-4 may improve the detection of DENV-4 cases worldwide. In addition, a serotype-specific NS1 Ag test identifies both DENV and the infecting serotype. Methods In this study, we used a small-ubiquitin-like modifier (SUMO*) cloning vector to express a SUMO*-DENV-4 rNS1 fusion protein to develop NS1 DENV-4 specific monoclonal antibodies (MAbs). These newly developed MAbs were then optimized for use in an anti-NS1 DENV-4 capture ELISA. The serotype specificity and sensitivity of this ELISA was evaluated using (i) supernatants from DENV (1–4)-infected Vero cell cultures, (ii) rNS1s from all the four DENV (1–4) and, (iii) rNS1s of related flaviViruses (yellow fever Virus; YFV and West Nile Virus; WNV). Results From the evaluation studies of the newly developed MAbs, we identified three DENV-4 specific anti-NS1 MAbs: 3H7A9, 8A6F2 and 6D4B10. Two of these MAbs were optimal for use in a DENV-4 serotype-specific NS1 capture ELISA: MAb 8A6F2 as the capture antibody and 6D4B10 as a detection antibody. Conclusion This ELISA was sensitive and specific to DENV-4 with no cross-reactivity to other three DENV (1–3) serotypes and other heterologous flaviViruses. Taken together these data indicated that our MAbs are useful reagents for the development of DENV-4 immunodiagnostic tests.

  • Development and characterization of serotype-specific monoclonal antibodies against the Dengue Virus-4 (DENV-4) non-structural protein (NS1).
    Virology journal, 2018
    Co-Authors: Tesfaye Gelanew, Elizabeth Hunsperger
    Abstract:

    Dengue, caused by one of the four serologically distinct Dengue Viruses (DENV-1 to − 4), is a mosquito-borne disease of serious global health significance. Reliable and cost-effective diagnostic tests, along with effective vaccines and vector-control strategies, are highly required to reduce Dengue morbidity and mortality. Evaluation studies revealed that many commercially available NS1 antigen (Ag) tests have limited sensitivity to DENV-4 serotype compared to the other three serotypes. These studies indicated the need for development of new NS1 Ag detection test with improved sensitivity to DENV-4. An NS1 capture enzyme linked immunoassay (ELISA) specific to DENV-4 may improve the detection of DENV-4 cases worldwide. In addition, a serotype-specific NS1 Ag test identifies both DENV and the infecting serotype. In this study, we used a small-ubiquitin-like modifier (SUMO*) cloning vector to express a SUMO*-DENV-4 rNS1 fusion protein to develop NS1 DENV-4 specific monoclonal antibodies (MAbs). These newly developed MAbs were then optimized for use in an anti-NS1 DENV-4 capture ELISA. The serotype specificity and sensitivity of this ELISA was evaluated using (i) supernatants from DENV (1–4)-infected Vero cell cultures, (ii) rNS1s from all the four DENV (1–4) and, (iii) rNS1s of related flaviViruses (yellow fever Virus; YFV and West Nile Virus; WNV). From the evaluation studies of the newly developed MAbs, we identified three DENV-4 specific anti-NS1 MAbs: 3H7A9, 8A6F2 and 6D4B10. Two of these MAbs were optimal for use in a DENV-4 serotype-specific NS1 capture ELISA: MAb 8A6F2 as the capture antibody and 6D4B10 as a detection antibody. This ELISA was sensitive and specific to DENV-4 with no cross-reactivity to other three DENV (1–3) serotypes and other heterologous flaviViruses. Taken together these data indicated that our MAbs are useful reagents for the development of DENV-4 immunodiagnostic tests.

Tesfaye Gelanew - One of the best experts on this subject based on the ideXlab platform.

  • Development and characterization of serotype-specific monoclonal antibodies against the Dengue Virus-4 (DENV-4) non-structural protein (NS1)
    Virology Journal, 2018
    Co-Authors: Tesfaye Gelanew, Elizabeth Hunsperger
    Abstract:

    Background Dengue, caused by one of the four serologically distinct Dengue Viruses (DENV-1 to − 4), is a mosquito-borne disease of serious global health significance. Reliable and cost-effective diagnostic tests, along with effective vaccines and vector-control strategies, are highly required to reduce Dengue morbidity and mortality. Evaluation studies revealed that many commercially available NS1 antigen (Ag) tests have limited sensitivity to DENV-4 serotype compared to the other three serotypes. These studies indicated the need for development of new NS1 Ag detection test with improved sensitivity to DENV-4. An NS1 capture enzyme linked immunoassay (ELISA) specific to DENV-4 may improve the detection of DENV-4 cases worldwide. In addition, a serotype-specific NS1 Ag test identifies both DENV and the infecting serotype. Methods In this study, we used a small-ubiquitin-like modifier (SUMO*) cloning vector to express a SUMO*-DENV-4 rNS1 fusion protein to develop NS1 DENV-4 specific monoclonal antibodies (MAbs). These newly developed MAbs were then optimized for use in an anti-NS1 DENV-4 capture ELISA. The serotype specificity and sensitivity of this ELISA was evaluated using (i) supernatants from DENV (1–4)-infected Vero cell cultures, (ii) rNS1s from all the four DENV (1–4) and, (iii) rNS1s of related flaviViruses (yellow fever Virus; YFV and West Nile Virus; WNV). Results From the evaluation studies of the newly developed MAbs, we identified three DENV-4 specific anti-NS1 MAbs: 3H7A9, 8A6F2 and 6D4B10. Two of these MAbs were optimal for use in a DENV-4 serotype-specific NS1 capture ELISA: MAb 8A6F2 as the capture antibody and 6D4B10 as a detection antibody. Conclusion This ELISA was sensitive and specific to DENV-4 with no cross-reactivity to other three DENV (1–3) serotypes and other heterologous flaviViruses. Taken together these data indicated that our MAbs are useful reagents for the development of DENV-4 immunodiagnostic tests.

  • Development and characterization of serotype-specific monoclonal antibodies against the Dengue Virus-4 (DENV-4) non-structural protein (NS1).
    Virology journal, 2018
    Co-Authors: Tesfaye Gelanew, Elizabeth Hunsperger
    Abstract:

    Dengue, caused by one of the four serologically distinct Dengue Viruses (DENV-1 to − 4), is a mosquito-borne disease of serious global health significance. Reliable and cost-effective diagnostic tests, along with effective vaccines and vector-control strategies, are highly required to reduce Dengue morbidity and mortality. Evaluation studies revealed that many commercially available NS1 antigen (Ag) tests have limited sensitivity to DENV-4 serotype compared to the other three serotypes. These studies indicated the need for development of new NS1 Ag detection test with improved sensitivity to DENV-4. An NS1 capture enzyme linked immunoassay (ELISA) specific to DENV-4 may improve the detection of DENV-4 cases worldwide. In addition, a serotype-specific NS1 Ag test identifies both DENV and the infecting serotype. In this study, we used a small-ubiquitin-like modifier (SUMO*) cloning vector to express a SUMO*-DENV-4 rNS1 fusion protein to develop NS1 DENV-4 specific monoclonal antibodies (MAbs). These newly developed MAbs were then optimized for use in an anti-NS1 DENV-4 capture ELISA. The serotype specificity and sensitivity of this ELISA was evaluated using (i) supernatants from DENV (1–4)-infected Vero cell cultures, (ii) rNS1s from all the four DENV (1–4) and, (iii) rNS1s of related flaviViruses (yellow fever Virus; YFV and West Nile Virus; WNV). From the evaluation studies of the newly developed MAbs, we identified three DENV-4 specific anti-NS1 MAbs: 3H7A9, 8A6F2 and 6D4B10. Two of these MAbs were optimal for use in a DENV-4 serotype-specific NS1 capture ELISA: MAb 8A6F2 as the capture antibody and 6D4B10 as a detection antibody. This ELISA was sensitive and specific to DENV-4 with no cross-reactivity to other three DENV (1–3) serotypes and other heterologous flaviViruses. Taken together these data indicated that our MAbs are useful reagents for the development of DENV-4 immunodiagnostic tests.

Nitwara Wikan - One of the best experts on this subject based on the ideXlab platform.

  • Delayed antibody dependent enhancement of low passage Dengue Virus 4 isolates
    BMC Research Notes, 2015
    Co-Authors: Nitwara Wikan, Prasert Auewarakul, Sirikwan Libsittikul, Sutee Yoksan, Duncan R. Smith
    Abstract:

    Background The concept of antibody dependent enhancement (ADE) of Dengue Virus (DENV) infection is a cornerstone of our current understanding of Dengue pathogenesis, although some questions as to the mechanism remain, particularly in regards to the behavior of low and high passage Virus isolates. This study utilized two low passage DENV 4 isolates and a laboratory adapted DENV 4 isolate to investigate the potential of low passage isolates to undergo ADE. Results Little or no ADE of infection was observed on day 2 post infection with low passage isolates, while high enhancement of infection was seen with the laboratory adapted Virus. However, both of the low passage isolates showed high levels of infection (60–100 %) by day 5 post infection. Conclusions These results show that low passage DENV 4 Viruses undergo ADE mediated infection, but that the process is significantly temporally delayed as compared to laboratory adapted DENV 4.

  • Delayed antibody dependent enhancement of low passage Dengue Virus 4 isolates
    BMC research notes, 2015
    Co-Authors: Nitwara Wikan, Prasert Auewarakul, Sirikwan Libsittikul, Sutee Yoksan, Duncan R. Smith
    Abstract:

    Background The concept of antibody dependent enhancement (ADE) of Dengue Virus (DENV) infection is a cornerstone of our current understanding of Dengue pathogenesis, although some questions as to the mechanism remain, particularly in regards to the behavior of low and high passage Virus isolates. This study utilized two low passage DENV 4 isolates and a laboratory adapted DENV 4 isolate to investigate the potential of low passage isolates to undergo ADE.

Prasert Auewarakul - One of the best experts on this subject based on the ideXlab platform.

  • Amino acid alignment of flaviVirus NS5 proteins.
    2018
    Co-Authors: Chonticha Saisawang, Atichat Kuadkitkan, Prasert Auewarakul, Duncan R. Smith, Albert J. Ketterman
    Abstract:

    The sequences are nonstructural protein NS5 from Dengue Virus 2 NP_739590.2, Dengue Virus 1 NP_722465.1, Dengue Virus 3 YP_001531176.2, Dengue Virus 4 NP_740325.1, Zika Virus AMR39834.1, Japanese encephalitis Virus NP_775674.1, West Nile Virus YP_001527887.1, Tick-borne encephalitis Virus NP_775511.1, Yellow fever Virus NP_776009.1, Murray Valley encephalitis Virus NP_722539.1, Langat Virus NP_740302.1. Cysteines are highlighted in yellow with the positions of the 3 glutathionylated cysteines identified in this study shown in green highlight in the alignment. Blue bars illustrate the residues of the finger subdomains, the red bars show the residues of the palm domain and the purple bars show the residues of the thumb domain [30]. Conservation of amino acid sequence is shown by 100 percent conserved as white on black, between 80 to 100 percent conserved as white on dark grey, between 60 to 80 percent conserved as black on light grey, and less than 60 percent conserved as black on white.

  • Delayed antibody dependent enhancement of low passage Dengue Virus 4 isolates
    BMC Research Notes, 2015
    Co-Authors: Nitwara Wikan, Prasert Auewarakul, Sirikwan Libsittikul, Sutee Yoksan, Duncan R. Smith
    Abstract:

    Background The concept of antibody dependent enhancement (ADE) of Dengue Virus (DENV) infection is a cornerstone of our current understanding of Dengue pathogenesis, although some questions as to the mechanism remain, particularly in regards to the behavior of low and high passage Virus isolates. This study utilized two low passage DENV 4 isolates and a laboratory adapted DENV 4 isolate to investigate the potential of low passage isolates to undergo ADE. Results Little or no ADE of infection was observed on day 2 post infection with low passage isolates, while high enhancement of infection was seen with the laboratory adapted Virus. However, both of the low passage isolates showed high levels of infection (60–100 %) by day 5 post infection. Conclusions These results show that low passage DENV 4 Viruses undergo ADE mediated infection, but that the process is significantly temporally delayed as compared to laboratory adapted DENV 4.

  • Delayed antibody dependent enhancement of low passage Dengue Virus 4 isolates
    BMC research notes, 2015
    Co-Authors: Nitwara Wikan, Prasert Auewarakul, Sirikwan Libsittikul, Sutee Yoksan, Duncan R. Smith
    Abstract:

    Background The concept of antibody dependent enhancement (ADE) of Dengue Virus (DENV) infection is a cornerstone of our current understanding of Dengue pathogenesis, although some questions as to the mechanism remain, particularly in regards to the behavior of low and high passage Virus isolates. This study utilized two low passage DENV 4 isolates and a laboratory adapted DENV 4 isolate to investigate the potential of low passage isolates to undergo ADE.