BACKGROUND: The objective of the study was to develop and evaluate IgM and IgG ELISAs and an IgG Western blot test for the serological detection of human infections with Andes virus (ANDV), the major cause of hantavirus cardiopulmonary syndrome (HCPS) in South America. METHODS: The entire nucleocapsid (N) protein-encoding sequence of ANDV (strain AH-1) was cloned and expressed in the yeast Saccharomyces cerevisiae. The polyhistidine-tagged recombinant N (rN) protein of ANDV was purified by nickel-chelation chromatography and characterized by its reactivity with different N-specific monoclonal antibodies. To detect an antibody response directed against ANDV in humans, indirect IgM and IgG ELISAs and an IgG Western blot test based on ANDV rN antigen were developed. The evaluation of the tests was performed using a negative serum panel and 63 blinded sera from Argentina and Chile, containing acute-phase and convalescent sera from HCPS patients. RESULTS: The specificities and sensitivities for the IgM and IgG ELISAs were demonstrated to be very high. The IgG ELISA data were confirmed by the IgG Western blot assay based on the same rN antigen. Almost all anti-ANDV-positive sera reacted to higher endpoint titers with N protein of ANDV than with those of Sin Nombre, Laguna Negra or Puumala virus. The cross-reactivity of anti-ANDV-N IgG-positive sera to rN proteins of other hantaviruses was found to be increased with time after the onset of HCPS. CONCLUSION: The high sensitivity of the novel assays should facilitate early diagnosis of ANDV infections and might contribute to a successful treatment of HCPS patients.
BACKGROUND: The objective of the study was to develop and evaluate IgM and IgG ELISAs and an IgG Western blot test for the serological detection of humaninfections with Andes virus (ANDV), the major cause of hantavirus cardiopulmonary syndrome (HCPS) in South America. METHODS: The entire nucleocapsid (N) protein-encoding sequence of ANDV (strain AH-1) was cloned and expressed in the yeastSaccharomyces cerevisiae. The polyhistidine-tagged recombinant N (rN) protein of ANDV was purified by nickel-chelation chromatography and characterized by its reactivity with different N-specific monoclonal antibodies. To detect an antibody response directed against ANDV in humans, indirect IgM and IgG ELISAs and an IgG Western blot test based on ANDV rN antigen were developed. The evaluation of the tests was performed using a negative serum panel and 63 blinded sera from Argentina and Chile, containing acute-phase and convalescent sera from HCPS patients. RESULTS: The specificities and sensitivities for the IgM and IgG ELISAs were demonstrated to be very high. The IgG ELISA data were confirmed by the IgG Western blot assay based on the same rN antigen. Almost all anti-ANDV-positive sera reacted to higher endpoint titers with N protein of ANDV than with those of Sin Nombre, Laguna Negra or Puumala virus. The cross-reactivity of anti-ANDV-N IgG-positive sera to rN proteins of other hantaviruses was found to be increased with time after the onset of HCPS. CONCLUSION: The high sensitivity of the novel assays should facilitate early diagnosis of ANDVinfections and might contribute to a successful treatment of HCPS patients.
Authors: Marc Mertens; Jörg Hofmann; Rasa Petraityte-Burneikiene; Mario Ziller; Kestutis Sasnauskas; Robert Friedrich; Olaf Niederstrasser; Detlev H Krüger; Martin H Groschup; Eckhardt Petri; Sandra Werdermann; Rainer G Ulrich Journal: Med Microbiol Immunol Date: 2011-05-25 Impact factor: 3.402
Authors: Marc Mertens; Eveline Kindler; Petra Emmerich; Jutta Esser; Christiane Wagner-Wiening; Roman Wölfel; Rasa Petraityte-Burneikiene; Jonas Schmidt-Chanasit; Aurelija Zvirbliene; Martin H Groschup; Gerhard Dobler; Martin Pfeffer; Gerald Heckel; Rainer G Ulrich; Sandra S Essbauer Journal: Virus Genes Date: 2011-05-20 Impact factor: 2.332
Authors: S Essbauer; J Schmidt; F J Conraths; R Friedrich; J Koch; W Hautmann; M Pfeffer; R Wölfel; J Finke; G Dobler; R Ulrich Journal: Epidemiol Infect Date: 2006-05-02 Impact factor: 2.451
Authors: Marc Mertens; Roman Wölfel; Katrin Ullrich; Kumiko Yoshimatsu; Jana Blumhardt; Ina Römer; Jutta Esser; Jonas Schmidt-Chanasit; Martin H Groschup; Gerhard Dobler; Sandra S Essbauer; Rainer G Ulrich Journal: Med Microbiol Immunol Date: 2009-01-16 Impact factor: 3.402
Authors: Sabine Lederer; Erik Lattwein; Merle Hanke; Karen Sonnenberg; Winfried Stoecker; Åke Lundkvist; Antti Vaheri; Olli Vapalahti; Paul K S Chan; Heinz Feldmann; Daryl Dick; Jonas Schmidt-Chanasit; Paula Padula; Pablo A Vial; Raluca Panculescu-Gatej; Cornelia Ceianu; Paul Heyman; Tatjana Avšič-Županc; Matthias Niedrig Journal: PLoS Negl Trop Dis Date: 2013-04-04