title: Development of a novel plaque reduction neutralisation test for hantavirus infection.
abstract: In the Americas, hantaviruses cause severe cardiopulmonary syndrome (HCPS) with a high fatality rate. Hantavirus infection is commonly diagnosed using serologic techniques and reverse transcription-polymerase chain reaction. This paper presents a novel plaque reduction neutralisation test (PRNT) for detecting antibodies to Brazilian hantavirus. Using PRNT, plaque detection was enhanced by adding 0.6% of dimethyl sulfoxide into the overlay culture medium of the infected cells. This procedure facilitated clear visualisation of small plaques under the microscope and provided for easy and accurate plaque counting. The sera from 37 HCPS patients from the city of Ribeirão Preto, Brazil was evaluated for the Rio Mamoré virus (RIOMV) using PRNT. Six samples exhibited neutralising antibodies; these antibodies exhibited a low titre. The low level of seropositive samples may be due to fewer cross-reactions between two different hantavirus species; the patients were likely infected by Araraquara virus (a virus that has not been isolated) and RIOMV was used for the test. This assay offers a new approach to evaluating and measuring neutralising antibodies produced during hantavirus infections and it can be adapted to other hantaviruses, including viruses that will be isolated in the future.
authors.affiliation: Centro de Pesquisa em Virologia, Faculdade de Medicina de Riberão Preto, Universidade de São Paulo, Ribeirão Preto, SP, BR.
authors.forename: Michelly de
authors.initials: Md
authors.lastname: Pádua
authors.affiliation: Centro de Pesquisa em Virologia, Faculdade de Medicina de Riberão Preto, Universidade de São Paulo, Ribeirão Preto, SP, BR.
authors.forename: William Marciel de
authors.initials: WM
authors.lastname: Souza
authors.affiliation: Centro de Pesquisa em Virologia, Faculdade de Medicina de Riberão Preto, Universidade de São Paulo, Ribeirão Preto, SP, BR.
authors.forename: Flávio
authors.initials: F
authors.lastname: Lauretti
authors.affiliation: Centro de Pesquisa em Virologia, Faculdade de Medicina de Riberão Preto, Universidade de São Paulo, Ribeirão Preto, SP, BR.
authors.forename: Luiz Tadeu Moraes
authors.initials: LT
authors.lastname: Figueiredo
mesh_terms: D000914:Antibodies, Viral / Q000097:blood* / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D006476:Orthohantavirus / Q000254:growth & development / Q000276:immunology; D018804:Hantavirus Pulmonary Syndrome / Q000175:diagnosis* / Q000821:virology; D006801:Humans; D009500:Neutralization Tests / Q000379:methods*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D010948:Viral Plaque Assay
chemical_list: D000914:Antibodies, Viral