Laboratory diagnosis of Toscana virus infection by enzyme immunoassay with recombinant viral nucleoprotein.
A recombinant enzyme immunoassay (rEIA) to detect serum immunoglobulin M (IgM) and IgG to Toscana virus (TOSV) was developed with the aim of establishing a simple and easily available assay for diagnosing acute and/or previous infections. The rEIA, based on the recombinant nucleoprotein of TOSV expressed in Escherichia coli, was evaluated with 97 serum samples collected in an area where TOSV is endemic and compared to an analogous assay based on cell-derived TOSV. Discordant results were resolved by immunoblotting (IB). Twenty-two of these samples, obtained from subjects hospitalized during the summer season with meningitis of suspected TOSV etiology, were further characterized by indirect immunofluorescence and IB, and detection of specific TOSV RNA sequences in the cerebrospinal fluid of these patients was attempted by nested PCR. The results indicated that rEIA was able to diagnose acute TOSV infection by detection of specific serum IgM in all of the subjects with TOSV meningitis confirmed by nested PCR or serology. The overall sensitivity and specificity of rEIA were both 100% for IgM detection and 100 and 96.6%, respectively, for IgG detection. Thus, rEIA appears to be a simple and reliable laboratory test for the diagnosis of acute TOSV infection and for the assessment of immune status.
Soldateschi
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D
DIESSE Diagnostica Senese Srl, 53035 Monteriggioni (SI), Italy. Diessi@tin.it
dal Maso
G M
GM
Valassina
M
M
Santini
L
L
Bianchi
S
S
Cusi
M G
MG
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D002522:Chlorocebus aethiops; D019411:Clinical Laboratory Techniques; D003001:Cloning, Molecular; D004926:Escherichia coli; D006801:Humans; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G / Q000097:blood; D007075:Immunoglobulin M / Q000097:blood; D019590:Nucleocapsid Proteins / Q000032:analysis* / Q000276:immunology; D010217:Phlebotomus Fever / Q000134:cerebrospinal fluid / Q000175:diagnosis* / Q000276:immunology; D016856:Phlebovirus / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000134:cerebrospinal fluid; D011994:Recombinant Proteins / Q000032:analysis / Q000276:immunology; D015203:Reproducibility of Results; D012621:Seasons; D012680:Sensitivity and Specificity; D014709:Vero Cells