title: Novel serological tools for detection of Thottapalayam virus, a Soricomorpha-borne hantavirus.
abstract: We developed serological tools for the detection of hantavirus-specific antibodies and hantavirus antigens in shrews. The work was focussed to generate Thottapalayam virus (TPMV)-specific monoclonal antibodies (mAbs) and anti-shrew immunoglobulin G (IgG) antibodies. The mAbs against TPMV nucleocapsid (N) protein were produced after immunization of BALB/c mice with recombinant TPMV N proteins expressed in Escherichia coli, baculovirus and Saccharomyces cerevisiae-mediated expression systems. In total, six TPMV N-protein-specific mAbs were generated that showed a characteristic fluorescent pattern in indirect immunofluorescence assay (IFA) using TPMV-infected Vero cells. Out of the six mAbs tested, five showed no cross-reaction to rodent-associated hantaviruses (Hantaan, Seoul, Puumala, Tula, Dobrava-Belgrade and Sin Nombre viruses) in IFA and enzyme-linked immunosorbent assay (ELISA), although one mAb reacted to Sin Nombre virus in IFA. None of the mAbs cross-reacted with an amino-terminal segment of the shrew-borne Asama virus N protein. Anti-shrew-IgG sera were prepared after immunization of rabbits and BALB/c-mice with protein-G-purified shrew IgG. TPMV-N-protein-specific sera were raised by immunisation of Asian house shrews (Suncus murinus) with purified yeast-expressed TPMV N protein. Using these tools, an indirect ELISA was developed to detect TPMV-N-protein-specific antibodies in the sera of shrews. Using an established serological assay, high TPMV N protein specific antibody titres were measured in the sera of TPMV-N-protein-immunized and experimentally TPMV-infected shrews, whereas no cross-reactivity to other hantavirus N proteins was found. Therefore, the generated mAbs and the established ELISA system represent useful serological tools to detect TPMV, TPMV-related virus antigens or hantavirus-specific antibodies in hantavirus-infected shrews.
authors: Friedrich-Loeffler-Institut, Institute for Novel and Emerging Infectious Diseases, Greifswald-Insel Riems, Germany.
authors: Mathias
authors: M
authors: Schlegel
authors: Erdenesaikhan
authors: E
authors: Tegshduuren
authors: Kumiko
authors: K
authors: Yoshimatsu
authors: Rasa
authors: R
authors: Petraityte
authors: Kestutis
authors: K
authors: Sasnauskas
authors: Bärbel
authors: B
authors: Hammerschmidt
authors: Robert
authors: R
authors: Friedrich
authors: Marc
authors: M
authors: Mertens
authors: Martin H
authors: MH
authors: Groschup
authors: Satoru
authors: S
authors: Arai
authors: Rika
authors: R
authors: Endo
authors: Kenta
authors: K
authors: Shimizu
authors: Takaaki
authors: T
authors: Koma
authors: Shumpei
authors: S
authors: Yasuda
authors: Chiaki
authors: C
authors: Ishihara
authors: Rainer G
authors: RG
authors: Ulrich
authors: Jiro
authors: J
authors: Arikawa
authors: Bernd
authors: B
authors: Köllner
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification; D000914:Antibodies, Viral / Q000302:isolation & purification; D000956:Antigens, Viral / Q000276:immunology; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D006476:Orthohantavirus / Q000145:classification / Q000302:isolation & purification*; D018778:Hantavirus Infections / Q000175:diagnosis / Q000662:veterinary*; D051379:Mice; D008807:Mice, Inbred BALB C; D019590:Nucleocapsid Proteins / Q000276:immunology; D011994:Recombinant Proteins / Q000276:immunology; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000379:methods; D012788:Shrews / Q000821:virology*; D014730:Veterinary Medicine / Q000379:methods; D014773:Virology / Q000379:methods
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D019590:Nucleocapsid Proteins; D011994:Recombinant Proteins