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EnglishBackground: Usutu virus (USUV), a flavivirus that belongs to the Japanese encephalitis virus (JEV) family, has recently emerged as a human pathogen, necessitating new diagnostic tools. Objective: The development and assessment of a real-time RT-PCR assay to detectUSUVinhumansamples. Study design: Based on USUV genomic sequences from GenBank, USUV-specific primers and probes that target the NS5 gene were designed. The sensitivity was evaluated in a 10-fold dilution series of plasmid that contained the amplicon and in a dilution series of a quantified human USUV isolate. The specificity was determined by testing various concentrations of related ArBo viruses, including flaviviruses and phleboviruses. Human RNAse P was also amplified in the assay. One hundred four human specimens from patients who suffered from viral meningoencephalitis were evaluated. Results: The real-time RT-PCR assay had a sensitivity of 50 genomic copies per reaction (corresponding to 2200 copies/ml) and 1 PFU/ml of USUV isolate. USUV isolates from Austria were identified with identical efficiency, and no ArBo viruses, other than USUV, were detected. USUV was also identified in 3 cerebrospinal fluid samples. All human samples were positive for RNAse P. Conclusions: This PCR assay is recommended for all cases in which a rapid and clinically accurate diagnosis of human USUV infection is required
| Titolo: | A rapid and specific real-time RT-PCR assay to identify Usutu virus in human plasma, serum, and cerebrospinal fluid |
| Autore/i: | CAVRINI, FRANCESCA; Pepa M. E.; GAIBANI, PAOLO; Pierro A; ROSSINI, GIADA; LANDINI, MARIA PAOLA; SAMBRI, VITTORIO |
| Autore/i Unibo: | |
| Anno: | 2011 |
| Rivista: | |
| Digital Object Identifier (DOI): | http://dx.doi.org/10.1016/j.jcv.2010.11.008 |
| Abstract: | Background: Usutu virus (USUV), a flavivirus that belongs to the Japanese encephalitis virus (JEV) family, has recently emerged as a human pathogen, necessitating new diagnostic tools. Objective: The development and assessment of a real-time RT-PCR assay to detectUSUVinhumansamples. Study design: Based on USUV genomic sequences from GenBank, USUV-specific primers and probes that target the NS5 gene were designed. The sensitivity was evaluated in a 10-fold dilution series of plasmid that contained the amplicon and in a dilution series of a quantified human USUV isolate. The specificity was determined by testing various concentrations of related ArBo viruses, including flaviviruses and phleboviruses. Human RNAse P was also amplified in the assay. One hundred four human specimens from patients who suffered from viral meningoencephalitis were evaluated. Results: The real-time RT-PCR assay had a sensitivity of 50 genomic copies per reaction (corresponding to 2200 copies/ml) and 1 PFU/ml of USUV isolate. USUV isolates from Austria were identified with identical efficiency, and no ArBo viruses, other than USUV, were detected. USUV was also identified in 3 cerebrospinal fluid samples. All human samples were positive for RNAse P. Conclusions: This PCR assay is recommended for all cases in which a rapid and clinically accurate diagnosis of human USUV infection is required |
| Data prodotto definitivo in UGOV: | 2013-06-13 09:06:28 |
| Appare nelle tipologie: | 1.01 Articolo in rivista |
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